JPH047200B2 - - Google Patents
Info
- Publication number
- JPH047200B2 JPH047200B2 JP4120283A JP4120283A JPH047200B2 JP H047200 B2 JPH047200 B2 JP H047200B2 JP 4120283 A JP4120283 A JP 4120283A JP 4120283 A JP4120283 A JP 4120283A JP H047200 B2 JPH047200 B2 JP H047200B2
- Authority
- JP
- Japan
- Prior art keywords
- nad
- reaction system
- dehydrogenase
- reaction
- adp
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired
Links
- 238000006243 chemical reaction Methods 0.000 claims description 100
- ZKHQWZAMYRWXGA-KQYNXXCUSA-J ATP(4-) Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP([O-])(=O)OP([O-])(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O ZKHQWZAMYRWXGA-KQYNXXCUSA-J 0.000 claims description 54
- ZKHQWZAMYRWXGA-UHFFFAOYSA-N Adenosine triphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(=O)OP(O)(O)=O)C(O)C1O ZKHQWZAMYRWXGA-UHFFFAOYSA-N 0.000 claims description 54
- DAYLJWODMCOQEW-TURQNECASA-N NMN zwitterion Chemical compound NC(=O)C1=CC=C[N+]([C@H]2[C@@H]([C@H](O)[C@@H](COP(O)([O-])=O)O2)O)=C1 DAYLJWODMCOQEW-TURQNECASA-N 0.000 claims description 45
- 239000000758 substrate Substances 0.000 claims description 45
- 238000000034 method Methods 0.000 claims description 40
- 239000005515 coenzyme Substances 0.000 claims description 39
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 27
- 230000001351 cycling effect Effects 0.000 claims description 26
- 102000004190 Enzymes Human genes 0.000 claims description 18
- 108090000790 Enzymes Proteins 0.000 claims description 18
- 125000003831 tetrazolyl group Chemical group 0.000 claims description 16
- 101710088194 Dehydrogenase Proteins 0.000 claims description 15
- 108091000080 Phosphotransferase Proteins 0.000 claims description 15
- 238000006911 enzymatic reaction Methods 0.000 claims description 15
- 102000020233 phosphotransferase Human genes 0.000 claims description 15
- 239000012085 test solution Substances 0.000 claims description 14
- 102000004316 Oxidoreductases Human genes 0.000 claims description 13
- 108090000854 Oxidoreductases Proteins 0.000 claims description 13
- -1 phosphorus compound Chemical class 0.000 claims description 13
- 102000005962 receptors Human genes 0.000 claims description 13
- 108020003175 receptors Proteins 0.000 claims description 13
- 102000007698 Alcohol dehydrogenase Human genes 0.000 claims description 12
- 108010021809 Alcohol dehydrogenase Proteins 0.000 claims description 12
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 12
- DRBBFCLWYRJSJZ-UHFFFAOYSA-N N-phosphocreatine Chemical compound OC(=O)CN(C)C(=N)NP(O)(O)=O DRBBFCLWYRJSJZ-UHFFFAOYSA-N 0.000 claims description 10
- 108010024137 Nicotinamide-Nucleotide Adenylyltransferase Proteins 0.000 claims description 10
- 102000015597 Nicotinamide-nucleotide adenylyltransferase Human genes 0.000 claims description 10
- 102000001253 Protein Kinase Human genes 0.000 claims description 10
- 235000011180 diphosphates Nutrition 0.000 claims description 10
- XPPKVPWEQAFLFU-UHFFFAOYSA-N diphosphoric acid Chemical compound OP(O)(=O)OP(O)(O)=O XPPKVPWEQAFLFU-UHFFFAOYSA-N 0.000 claims description 10
- 238000000691 measurement method Methods 0.000 claims description 10
- 238000006479 redox reaction Methods 0.000 claims description 9
- 102000000587 Glycerolphosphate Dehydrogenase Human genes 0.000 claims description 7
- 108010041921 Glycerolphosphate Dehydrogenase Proteins 0.000 claims description 7
- LIPOUNRJVLNBCD-UHFFFAOYSA-N acetyl dihydrogen phosphate Chemical compound CC(=O)OP(O)(O)=O LIPOUNRJVLNBCD-UHFFFAOYSA-N 0.000 claims description 7
- 229910052698 phosphorus Inorganic materials 0.000 claims description 7
- 239000011574 phosphorus Substances 0.000 claims description 7
- 108010052832 Cytochromes Proteins 0.000 claims description 6
- 102000018832 Cytochromes Human genes 0.000 claims description 6
- 101000892220 Geobacillus thermodenitrificans (strain NG80-2) Long-chain-alcohol dehydrogenase 1 Proteins 0.000 claims description 6
- RXKJFZQQPQGTFL-UHFFFAOYSA-N dihydroxyacetone Chemical compound OCC(=O)CO RXKJFZQQPQGTFL-UHFFFAOYSA-N 0.000 claims description 6
- 102000004420 Creatine Kinase Human genes 0.000 claims description 5
- 108010042126 Creatine kinase Proteins 0.000 claims description 5
- 102000003855 L-lactate dehydrogenase Human genes 0.000 claims description 5
- 108700023483 L-lactate dehydrogenases Proteins 0.000 claims description 5
- 102000013460 Malate Dehydrogenase Human genes 0.000 claims description 5
- 108010026217 Malate Dehydrogenase Proteins 0.000 claims description 5
- 239000002777 nucleoside Substances 0.000 claims description 5
- 229930029653 phosphoenolpyruvate Natural products 0.000 claims description 5
- DTBNBXWJWCWCIK-UHFFFAOYSA-N phosphoenolpyruvic acid Chemical compound OC(=O)C(=C)OP(O)(O)=O DTBNBXWJWCWCIK-UHFFFAOYSA-N 0.000 claims description 5
- 108060006633 protein kinase Proteins 0.000 claims description 5
- 150000004053 quinones Chemical class 0.000 claims description 5
- 239000004094 surface-active agent Substances 0.000 claims description 5
- 101710172561 3alpha-hydroxysteroid dehydrogenase Proteins 0.000 claims description 4
- PQGCEDQWHSBAJP-TXICZTDVSA-N 5-O-phosphono-alpha-D-ribofuranosyl diphosphate Chemical compound O[C@H]1[C@@H](O)[C@@H](O[P@](O)(=O)OP(O)(O)=O)O[C@@H]1COP(O)(O)=O PQGCEDQWHSBAJP-TXICZTDVSA-N 0.000 claims description 4
- 108010092060 Acetate kinase Proteins 0.000 claims description 4
- 102100036504 Dehydrogenase/reductase SDR family member 9 Human genes 0.000 claims description 4
- 108091006149 Electron carriers Proteins 0.000 claims description 4
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 claims description 4
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 claims description 4
- 150000002211 flavins Chemical class 0.000 claims description 4
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 claims description 3
- IXZNKTPIYKDIGG-REOHCLBHSA-L 4-phosphonato-L-aspartic acid(2-) Chemical compound [O-]C(=O)[C@@H]([NH3+])CC(=O)OP([O-])([O-])=O IXZNKTPIYKDIGG-REOHCLBHSA-L 0.000 claims description 3
- 102000002281 Adenylate kinase Human genes 0.000 claims description 3
- 108020000543 Adenylate kinase Proteins 0.000 claims description 3
- 108010020366 Arginine kinase Proteins 0.000 claims description 3
- 101000950981 Bacillus subtilis (strain 168) Catabolic NAD-specific glutamate dehydrogenase RocG Proteins 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- 108010050375 Glucose 1-Dehydrogenase Proteins 0.000 claims description 3
- 102000016901 Glutamate dehydrogenase Human genes 0.000 claims description 3
- 229930195714 L-glutamate Natural products 0.000 claims description 3
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 claims description 3
- JVTAAEKCZFNVCJ-REOHCLBHSA-N L-lactic acid Chemical compound C[C@H](O)C(O)=O JVTAAEKCZFNVCJ-REOHCLBHSA-N 0.000 claims description 3
- 108010064862 Nicotinamide phosphoribosyltransferase Proteins 0.000 claims description 3
- 102000015532 Nicotinamide phosphoribosyltransferase Human genes 0.000 claims description 3
- 102000011755 Phosphoglycerate Kinase Human genes 0.000 claims description 3
- 102000013009 Pyruvate Kinase Human genes 0.000 claims description 3
- 108020005115 Pyruvate Kinase Proteins 0.000 claims description 3
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 3
- 229940120503 dihydroxyacetone Drugs 0.000 claims description 3
- 239000008103 glucose Substances 0.000 claims description 3
- 229940116871 l-lactate Drugs 0.000 claims description 3
- 229910052760 oxygen Inorganic materials 0.000 claims description 3
- 239000001301 oxygen Substances 0.000 claims description 3
- CCTIOCVIZPCTGO-BYPYZUCNSA-N phosphoarginine Chemical compound OC(=O)[C@@H](N)CCCNC(=N)NP(O)(O)=O CCTIOCVIZPCTGO-BYPYZUCNSA-N 0.000 claims description 3
- AWUCVROLDVIAJX-GSVOUGTGSA-N sn-glycerol 3-phosphate Chemical compound OC[C@@H](O)COP(O)(O)=O AWUCVROLDVIAJX-GSVOUGTGSA-N 0.000 claims description 3
- CCBICDLNWJRFPO-UHFFFAOYSA-N 2,6-dichloroindophenol Chemical compound C1=CC(O)=CC=C1N=C1C=C(Cl)C(=O)C(Cl)=C1 CCBICDLNWJRFPO-UHFFFAOYSA-N 0.000 claims description 2
- 108010055400 Aspartate kinase Proteins 0.000 claims description 2
- 108020004827 Carbamate kinase Proteins 0.000 claims description 2
- JLVVSXFLKOJNIY-UHFFFAOYSA-N Magnesium ion Chemical compound [Mg+2] JLVVSXFLKOJNIY-UHFFFAOYSA-N 0.000 claims description 2
- 101001099217 Thermotoga maritima (strain ATCC 43589 / DSM 3109 / JCM 10099 / NBRC 100826 / MSB8) Triosephosphate isomerase Proteins 0.000 claims description 2
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 claims description 2
- TTWYZDPBDWHJOR-IDIVVRGQSA-L adenosine triphosphate disodium Chemical compound [Na+].[Na+].C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(=O)OP([O-])([O-])=O)[C@@H](O)[C@H]1O TTWYZDPBDWHJOR-IDIVVRGQSA-L 0.000 claims description 2
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 claims description 2
- FFQKYPRQEYGKAF-UHFFFAOYSA-N carbamoyl phosphate Chemical compound NC(=O)OP(O)(O)=O FFQKYPRQEYGKAF-UHFFFAOYSA-N 0.000 claims description 2
- 239000001177 diphosphate Substances 0.000 claims description 2
- YAGKRVSRTSUGEY-UHFFFAOYSA-N ferricyanide Chemical class [Fe+3].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-].N#[C-] YAGKRVSRTSUGEY-UHFFFAOYSA-N 0.000 claims description 2
- 229910001425 magnesium ion Inorganic materials 0.000 claims description 2
- 229960003966 nicotinamide Drugs 0.000 claims description 2
- 235000005152 nicotinamide Nutrition 0.000 claims description 2
- 239000011570 nicotinamide Substances 0.000 claims description 2
- 239000002736 nonionic surfactant Substances 0.000 claims description 2
- YNCMLFHHXWETLD-UHFFFAOYSA-N pyocyanin Chemical compound CN1C2=CC=CC=C2N=C2C1=CC=CC2=O YNCMLFHHXWETLD-UHFFFAOYSA-N 0.000 claims description 2
- OIRDTQYFTABQOQ-KQYNXXCUSA-N adenosine Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](CO)[C@@H](O)[C@H]1O OIRDTQYFTABQOQ-KQYNXXCUSA-N 0.000 claims 2
- RBNPOMFGQQGHHO-UHFFFAOYSA-N -2,3-Dihydroxypropanoic acid Natural products OCC(O)C(O)=O RBNPOMFGQQGHHO-UHFFFAOYSA-N 0.000 claims 1
- 239000002126 C01EB10 - Adenosine Substances 0.000 claims 1
- 229960005305 adenosine Drugs 0.000 claims 1
- 150000002148 esters Chemical class 0.000 claims 1
- 239000003153 chemical reaction reagent Substances 0.000 description 11
- 238000005259 measurement Methods 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 239000011777 magnesium Substances 0.000 description 8
- 238000002835 absorbance Methods 0.000 description 7
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 7
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 238000011088 calibration curve Methods 0.000 description 6
- 229940048084 pyrophosphate Drugs 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- 239000000126 substance Substances 0.000 description 6
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 5
- AZQWKYJCGOJGHM-UHFFFAOYSA-N 1,4-benzoquinone Chemical compound O=C1C=CC(=O)C=C1 AZQWKYJCGOJGHM-UHFFFAOYSA-N 0.000 description 4
- 239000002253 acid Substances 0.000 description 4
- GNGACRATGGDKBX-UHFFFAOYSA-N dihydroxyacetone phosphate Chemical compound OCC(=O)COP(O)(O)=O GNGACRATGGDKBX-UHFFFAOYSA-N 0.000 description 4
- 239000007788 liquid Substances 0.000 description 4
- 238000011002 quantification Methods 0.000 description 4
- 238000006722 reduction reaction Methods 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 241000194107 Bacillus megaterium Species 0.000 description 3
- 238000010521 absorption reaction Methods 0.000 description 3
- 230000009471 action Effects 0.000 description 3
- 239000003513 alkali Substances 0.000 description 3
- 230000003321 amplification Effects 0.000 description 3
- 239000012491 analyte Substances 0.000 description 3
- 239000000872 buffer Substances 0.000 description 3
- 238000006386 neutralization reaction Methods 0.000 description 3
- 238000003199 nucleic acid amplification method Methods 0.000 description 3
- 230000035484 reaction time Effects 0.000 description 3
- 230000009467 reduction Effects 0.000 description 3
- FRASJONUBLZVQX-UHFFFAOYSA-N 1,4-naphthoquinone Chemical compound C1=CC=C2C(=O)C=CC(=O)C2=C1 FRASJONUBLZVQX-UHFFFAOYSA-N 0.000 description 2
- OSJPPGNTCRNQQC-UWTATZPHSA-N 3-phospho-D-glyceric acid Chemical compound OC(=O)[C@H](O)COP(O)(O)=O OSJPPGNTCRNQQC-UWTATZPHSA-N 0.000 description 2
- XTWYTFMLZFPYCI-KQYNXXCUSA-N 5'-adenylphosphoric acid Chemical compound C1=NC=2C(N)=NC=NC=2N1[C@@H]1O[C@H](COP(O)(=O)OP(O)(O)=O)[C@@H](O)[C@H]1O XTWYTFMLZFPYCI-KQYNXXCUSA-N 0.000 description 2
- IKHGUXGNUITLKF-UHFFFAOYSA-N Acetaldehyde Chemical compound CC=O IKHGUXGNUITLKF-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- MJVAVZPDRWSRRC-UHFFFAOYSA-N Menadione Chemical compound C1=CC=C2C(=O)C(C)=CC(=O)C2=C1 MJVAVZPDRWSRRC-UHFFFAOYSA-N 0.000 description 2
- 102000002247 NADPH Dehydrogenase Human genes 0.000 description 2
- 108010014870 NADPH Dehydrogenase Proteins 0.000 description 2
- ACFIXJIJDZMPPO-NNYOXOHSSA-J NADPH(4-) Chemical compound C1=CCC(C(=O)N)=CN1[C@H]1[C@H](O)[C@H](O)[C@@H](COP([O-])(=O)OP([O-])(=O)OC[C@@H]2[C@H]([C@@H](OP([O-])([O-])=O)[C@@H](O2)N2C3=NC=NC(N)=C3N=C2)O)O1 ACFIXJIJDZMPPO-NNYOXOHSSA-J 0.000 description 2
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 description 2
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 2
- DFPAKSUCGFBDDF-ZQBYOMGUSA-N [14c]-nicotinamide Chemical compound N[14C](=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-ZQBYOMGUSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- CVSVTCORWBXHQV-UHFFFAOYSA-N creatine Chemical compound NC(=[NH2+])N(C)CC([O-])=O CVSVTCORWBXHQV-UHFFFAOYSA-N 0.000 description 2
- 230000002255 enzymatic effect Effects 0.000 description 2
- VWWQXMAJTJZDQX-UYBVJOGSSA-N flavin adenine dinucleotide Chemical compound C1=NC2=C(N)N=CN=C2N1[C@@H]([C@H](O)[C@@H]1O)O[C@@H]1CO[P@](O)(=O)O[P@@](O)(=O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C2=NC(=O)NC(=O)C2=NC2=C1C=C(C)C(C)=C2 VWWQXMAJTJZDQX-UYBVJOGSSA-N 0.000 description 2
- 235000019162 flavin adenine dinucleotide Nutrition 0.000 description 2
- 239000011714 flavin adenine dinucleotide Substances 0.000 description 2
- FVTCRASFADXXNN-SCRDCRAPSA-N flavin mononucleotide Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-SCRDCRAPSA-N 0.000 description 2
- 239000011768 flavin mononucleotide Substances 0.000 description 2
- 229940013640 flavin mononucleotide Drugs 0.000 description 2
- FVTCRASFADXXNN-UHFFFAOYSA-N flavin mononucleotide Natural products OP(=O)(O)OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O FVTCRASFADXXNN-UHFFFAOYSA-N 0.000 description 2
- 229940093632 flavin-adenine dinucleotide Drugs 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- CXKWCBBOMKCUKX-UHFFFAOYSA-M methylene blue Chemical compound [Cl-].C1=CC(N(C)C)=CC2=[S+]C3=CC(N(C)C)=CC=C3N=C21 CXKWCBBOMKCUKX-UHFFFAOYSA-M 0.000 description 2
- 229960000907 methylthioninium chloride Drugs 0.000 description 2
- 230000003647 oxidation Effects 0.000 description 2
- 238000007254 oxidation reaction Methods 0.000 description 2
- 239000008363 phosphate buffer Substances 0.000 description 2
- 150000003018 phosphorus compounds Chemical class 0.000 description 2
- 229940005657 pyrophosphoric acid Drugs 0.000 description 2
- 235000019231 riboflavin-5'-phosphate Nutrition 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- GPRLSGONYQIRFK-MNYXATJNSA-N triton Chemical compound [3H+] GPRLSGONYQIRFK-MNYXATJNSA-N 0.000 description 2
- UIXPTCZPFCVOQF-UHFFFAOYSA-N ubiquinone-0 Chemical compound COC1=C(OC)C(=O)C(C)=CC1=O UIXPTCZPFCVOQF-UHFFFAOYSA-N 0.000 description 2
- KPGXRSRHYNQIFN-UHFFFAOYSA-N 2-oxoglutaric acid Chemical compound OC(=O)CCC(=O)C(O)=O KPGXRSRHYNQIFN-UHFFFAOYSA-N 0.000 description 1
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- XTWYTFMLZFPYCI-UHFFFAOYSA-N Adenosine diphosphate Natural products C1=NC=2C(N)=NC=NC=2N1C1OC(COP(O)(=O)OP(O)(O)=O)C(O)C1O XTWYTFMLZFPYCI-UHFFFAOYSA-N 0.000 description 1
- ACTIUHUUMQJHFO-UHFFFAOYSA-N Coenzym Q10 Natural products COC1=C(OC)C(=O)C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UHFFFAOYSA-N 0.000 description 1
- 102100025287 Cytochrome b Human genes 0.000 description 1
- 108010075028 Cytochromes b Proteins 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- PHOQVHQSTUBQQK-SQOUGZDYSA-N D-glucono-1,5-lactone Chemical compound OC[C@H]1OC(=O)[C@H](O)[C@@H](O)[C@@H]1O PHOQVHQSTUBQQK-SQOUGZDYSA-N 0.000 description 1
- 108020005199 Dehydrogenases Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 102000005548 Hexokinase Human genes 0.000 description 1
- 108700040460 Hexokinases Proteins 0.000 description 1
- XUJNEKJLAYXESH-REOHCLBHSA-N L-Cysteine Chemical compound SC[C@H](N)C(O)=O XUJNEKJLAYXESH-REOHCLBHSA-N 0.000 description 1
- PWKSKIMOESPYIA-BYPYZUCNSA-N L-N-acetyl-Cysteine Chemical compound CC(=O)N[C@@H](CS)C(O)=O PWKSKIMOESPYIA-BYPYZUCNSA-N 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-N L-arginine Chemical compound OC(=O)[C@@H](N)CCCN=C(N)N ODKSFYDXXFIFQN-BYPYZUCNSA-N 0.000 description 1
- 229930064664 L-arginine Natural products 0.000 description 1
- 235000014852 L-arginine Nutrition 0.000 description 1
- CKLJMWTZIZZHCS-REOHCLBHSA-N L-aspartic acid Chemical compound OC(=O)[C@@H](N)CC(O)=O CKLJMWTZIZZHCS-REOHCLBHSA-N 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 229930192627 Naphthoquinone Natural products 0.000 description 1
- 108090000913 Nitrate Reductases Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 108700023219 Phosphoglycerate kinases Proteins 0.000 description 1
- 108010042687 Pyruvate Oxidase Proteins 0.000 description 1
- QAOWNCQODCNURD-UHFFFAOYSA-L Sulfate Chemical compound [O-]S([O-])(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-L 0.000 description 1
- 239000013504 Triton X-100 Substances 0.000 description 1
- 229920004890 Triton X-100 Polymers 0.000 description 1
- YMOPVQQBWLGDOD-UUOKFMHZSA-N XDP Chemical compound O[C@@H]1[C@H](O)[C@@H](COP(O)(=O)OP(O)(O)=O)O[C@H]1N1C(NC(=O)NC2=O)=C2N=C1 YMOPVQQBWLGDOD-UUOKFMHZSA-N 0.000 description 1
- XJLXINKUBYWONI-DQQFMEOOSA-N [[(2r,3r,4r,5r)-5-(6-aminopurin-9-yl)-3-hydroxy-4-phosphonooxyoxolan-2-yl]methoxy-hydroxyphosphoryl] [(2s,3r,4s,5s)-5-(3-carbamoylpyridin-1-ium-1-yl)-3,4-dihydroxyoxolan-2-yl]methyl phosphate Chemical compound NC(=O)C1=CC=C[N+]([C@@H]2[C@H]([C@@H](O)[C@H](COP([O-])(=O)OP(O)(=O)OC[C@@H]3[C@H]([C@@H](OP(O)(O)=O)[C@@H](O3)N3C4=NC=NC(N)=C4N=C3)O)O2)O)=C1 XJLXINKUBYWONI-DQQFMEOOSA-N 0.000 description 1
- 229960004308 acetylcysteine Drugs 0.000 description 1
- 238000010306 acid treatment Methods 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 150000004054 benzoquinones Chemical class 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 239000012295 chemical reaction liquid Substances 0.000 description 1
- 239000003638 chemical reducing agent Substances 0.000 description 1
- ACTIUHUUMQJHFO-UPTCCGCDSA-N coenzyme Q10 Chemical compound COC1=C(OC)C(=O)C(C\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CC\C=C(/C)CCC=C(C)C)=C(C)C1=O ACTIUHUUMQJHFO-UPTCCGCDSA-N 0.000 description 1
- 235000017471 coenzyme Q10 Nutrition 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 229960003624 creatine Drugs 0.000 description 1
- 239000006046 creatine Substances 0.000 description 1
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 description 1
- 235000018417 cysteine Nutrition 0.000 description 1
- 229960002433 cysteine Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 150000002019 disulfides Chemical class 0.000 description 1
- VHJLVAABSRFDPM-QWWZWVQMSA-N dithiothreitol Chemical compound SC[C@@H](O)[C@H](O)CS VHJLVAABSRFDPM-QWWZWVQMSA-N 0.000 description 1
- WAMKWBHYPYBEJY-UHFFFAOYSA-N duroquinone Chemical compound CC1=C(C)C(=O)C(C)=C(C)C1=O WAMKWBHYPYBEJY-UHFFFAOYSA-N 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 235000012209 glucono delta-lactone Nutrition 0.000 description 1
- 239000000182 glucono-delta-lactone Substances 0.000 description 1
- 229960003681 gluconolactone Drugs 0.000 description 1
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 229910052751 metal Inorganic materials 0.000 description 1
- 239000002184 metal Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 150000002791 naphthoquinones Chemical class 0.000 description 1
- FSVCQIDHPKZJSO-UHFFFAOYSA-L nitro blue tetrazolium dichloride Chemical compound [Cl-].[Cl-].COC1=CC(C=2C=C(OC)C(=CC=2)[N+]=2N(N=C(N=2)C=2C=CC=CC=2)C=2C=CC(=CC=2)[N+]([O-])=O)=CC=C1[N+]1=NC(C=2C=CC=CC=2)=NN1C1=CC=C([N+]([O-])=O)C=C1 FSVCQIDHPKZJSO-UHFFFAOYSA-L 0.000 description 1
- 210000004940 nucleus Anatomy 0.000 description 1
- 239000003129 oil well Substances 0.000 description 1
- KHPXUQMNIQBQEV-UHFFFAOYSA-N oxaloacetic acid Chemical compound OC(=O)CC(=O)C(O)=O KHPXUQMNIQBQEV-UHFFFAOYSA-N 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 125000002467 phosphate group Chemical group [H]OP(=O)(O[H])O[*] 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- NPCOQXAVBJJZBQ-UHFFFAOYSA-N reduced coenzyme Q9 Natural products COC1=C(O)C(C)=C(CC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)CCC=C(C)C)C(O)=C1OC NPCOQXAVBJJZBQ-UHFFFAOYSA-N 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 235000019192 riboflavin Nutrition 0.000 description 1
- 239000002151 riboflavin Substances 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 229940035936 ubiquinone Drugs 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- DGVVWUTYPXICAM-UHFFFAOYSA-N βâMercaptoethanol Chemical compound OCCS DGVVWUTYPXICAM-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Description
ãçºæã®è©³çްãªèª¬æã
æ¬çºæã¯ãATPïŒã¢ããã·ã³ããªãã¹ããšãŒ
ãïŒãŸãã¯NMNïŒãã³ãã³é žã¢ããã¢ããã¯ã¬
ãªããïŒã®é«æåºŠæž¬å®æ³ã«é¢ãããæŽã«è©³ãã
ã¯ãATPããã³NMNã®ããããäžæåã嫿
ããè¢«æ€æ¶²ã«ãäž»åå¿ç³»ã«ãããŠNMNãŸãã¯
ATPãMg2+ïŒãã°ãã·ãŠã ã€ãªã³ïŒã®ååšäžã«
NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒãäœçšãã
ãŠNADãšãããªã³é žã«å€æãããäž»åå¿ã®åŸã
çããNADããNADïŒïŒ°ïŒãè£é µçŽ ãšããé žå
éå åå¿ç³»ãããšãã°å°ãªããšãNADãæ¶è²»ã
ãŠéå åNADãçæããåå¿ã圢æããããã
ãã²ããŒãŒããã³ãã®åºè³ªã«ããåå¿ç³»ãšãéå
åNADïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»ããš
ãã°éå åNADãæ¶è²»ããŠNADãçæãããžã¢
ãã©ãŒãŒããã³ããã©ãŸãªãŠã å¡©ãšã«ããåå¿ç³»
ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯ãªã³ã°åå¿ãè¡ãª
ããæ¬¡ãã§åå¿ã«ãããŠçæãŸãã¯æ¶è²»ãããæ
åãå®éããŠãªãATPããã³NMNã®ãããã
äžæåã嫿ããè¢«æ€æ¶²äžã®æåã®é«æåºŠæž¬å®æ³
ã«é¢ããã
ãïŒãŸãã¯NMNïŒãã³ãã³é žã¢ããã¢ããã¯ã¬
ãªããïŒã®é«æåºŠæž¬å®æ³ã«é¢ãããæŽã«è©³ãã
ã¯ãATPããã³NMNã®ããããäžæåã嫿
ããè¢«æ€æ¶²ã«ãäž»åå¿ç³»ã«ãããŠNMNãŸãã¯
ATPãMg2+ïŒãã°ãã·ãŠã ã€ãªã³ïŒã®ååšäžã«
NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒãäœçšãã
ãŠNADãšãããªã³é žã«å€æãããäž»åå¿ã®åŸã
çããNADããNADïŒïŒ°ïŒãè£é µçŽ ãšããé žå
éå åå¿ç³»ãããšãã°å°ãªããšãNADãæ¶è²»ã
ãŠéå åNADãçæããåå¿ã圢æããããã
ãã²ããŒãŒããã³ãã®åºè³ªã«ããåå¿ç³»ãšãéå
åNADïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»ããš
ãã°éå åNADãæ¶è²»ããŠNADãçæãããžã¢
ãã©ãŒãŒããã³ããã©ãŸãªãŠã å¡©ãšã«ããåå¿ç³»
ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯ãªã³ã°åå¿ãè¡ãª
ããæ¬¡ãã§åå¿ã«ãããŠçæãŸãã¯æ¶è²»ãããæ
åãå®éããŠãªãATPããã³NMNã®ãããã
äžæåã嫿ããè¢«æ€æ¶²äžã®æåã®é«æåºŠæž¬å®æ³
ã«é¢ããã
NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒ
ïŒEC.2.7.7.1ïŒã¯äžèšã®é µçŽ äœçšãæããããºãã®
èèãè³çްèãããã³ãã¿ã®èèã®çŽ°èæ žã«ååš
ããããšãç¥ãããŠããïŒM.R.AtkinsonãJ.F.
JacksonãR.K.MortonãBiochem.J.ã80318
ïŒ1961ïŒïŒã
ïŒEC.2.7.7.1ïŒã¯äžèšã®é µçŽ äœçšãæããããºãã®
èèãè³çްèãããã³ãã¿ã®èèã®çŽ°èæ žã«ååš
ããããšãç¥ãããŠããïŒM.R.AtkinsonãJ.F.
JacksonãR.K.MortonãBiochem.J.ã80318
ïŒ1961ïŒïŒã
ATPïŒNMNMg2
ââââ
ãNADïŒãããªã³é
ž
æ¬é
µçŽ ã®æŽ»æ§æž¬å®æ³ãšããŠã¯ãåå¿ã«ããçã
ãNADãã¢ã«ã³ãŒã«ããããã²ããŒãŒ
ïŒEC.1.1.1.1ïŒã§éå ããŠãçããéå åNADã
340nïœã«ããåžå 床枬å®ããæ¹æ³ã®å ±åããªã
ããŠããïŒA.KornbergãâMethods in
Enzymologyâãvol.ãP.670ïŒ1955ïŒããé µçŽ ãã³
ãããã¯ãP.386ïŒ1982.12.1ïŒïŒããããããã®æŽ»æ§
æž¬å®æ³ã«åºããŠATPãŸãã¯NMNã®å®éæ³ãšãª
ããŠãããã®æåºŠãäœããATPãŸãã¯NMNã®
å®éæ³ãšããŠã¯å©çšãé£ããã®ã§ãã€ãã
ãNADãã¢ã«ã³ãŒã«ããããã²ããŒãŒ
ïŒEC.1.1.1.1ïŒã§éå ããŠãçããéå åNADã
340nïœã«ããåžå 床枬å®ããæ¹æ³ã®å ±åããªã
ããŠããïŒA.KornbergãâMethods in
Enzymologyâãvol.ãP.670ïŒ1955ïŒããé µçŽ ãã³
ãããã¯ãP.386ïŒ1982.12.1ïŒïŒããããããã®æŽ»æ§
æž¬å®æ³ã«åºããŠATPãŸãã¯NMNã®å®éæ³ãšãª
ããŠãããã®æåºŠãäœããATPãŸãã¯NMNã®
å®éæ³ãšããŠã¯å©çšãé£ããã®ã§ãã€ãã
æ¬çºæè
ã¯ãäžèšåå¿ã«ããçããNADãæž¬
å®ããããšã«ãããããã«ãã®NADããNAD
ïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»ãšéå å
NADïŒïŒ°ïŒãè£é µçŽ ãšããåå¿ç³»ãšã®çµåãã«ã
ãè£é µçŽ ãµã€ã¯ãªã³ã°åå¿ã«ããå¢å¹ åå¿ããã
çæãŸãã¯æ¶è²»ãããæåãå®éããããšã«ãã
é«æåºŠã«æž¬å®ã§ããããšãèŠãåºããã
å®ããããšã«ãããããã«ãã®NADããNAD
ïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»ãšéå å
NADïŒïŒ°ïŒãè£é µçŽ ãšããåå¿ç³»ãšã®çµåãã«ã
ãè£é µçŽ ãµã€ã¯ãªã³ã°åå¿ã«ããå¢å¹ åå¿ããã
çæãŸãã¯æ¶è²»ãããæåãå®éããããšã«ãã
é«æåºŠã«æž¬å®ã§ããããšãèŠãåºããã
è£é
µçŽ ãµã€ã¯ãªã³ã°ã¯ãåŸæ¥ããããç¥ããã
æ¹æ³ã§ããïŒçååŠå®éšè¬åº§ïŒãé µçŽ ç ç©¶æ³ãäž
121ã135ãæ¥æ¬çååŠäŒç·šãæ±äº¬ååŠå人ïŒãåº
質ãé µçŽ æŽ»æ§ãå¢å¹ å®éããæ¹æ³ã§ãããçäœå
ã®åŸ®éæåã®æž¬å®ã«ã¯é©ããæ¹æ³ã§ããã
æ¹æ³ã§ããïŒçååŠå®éšè¬åº§ïŒãé µçŽ ç ç©¶æ³ãäž
121ã135ãæ¥æ¬çååŠäŒç·šãæ±äº¬ååŠå人ïŒãåº
質ãé µçŽ æŽ»æ§ãå¢å¹ å®éããæ¹æ³ã§ãããçäœå
ã®åŸ®éæåã®æž¬å®ã«ã¯é©ããæ¹æ³ã§ããã
ããããªãããåŸæ¥ãããªãããŠãããµã€ã¯ãª
ã³ã°æ³ã§ã¯ãäž»åå¿ç³»ã«ã¯NADããã³éå å
NADã®äž¡æ¹ãå ±åããŠããããã®NADããã³é
å åNADã®ããããäžæ¹ãå®éããå Žåã«ã¯ã
ãŸãåå¿ç³»ã«å ±åããå®é察象å€ã®NADãŸãã¯
éå åNADã®ãããããæ¶å»ããã¢ã«ã«ãªãŸã
ã¯é žã®æ·»å ãå ç±åŠçã®æäœãå¿ é ãšããæ¬¡ãã§
PHããµã€ã¯ãªã³ã°åå¿ã«é©ããå€ã«èª¿æŽåŸãµã€ã¯
ãªã³ã°åå¿ãè¡ãªããããã«æç€ºåå¿ããããªã
ãªããã°ãªãããæäœæ§ã«åé¡ãããããã®æç€º
åå¿èªäœã被枬å®ç©è³ªä»¥å€ã®åœ±é¿ãåããå Žåã
å€ãã€ãããããæ²¹äºã®ãããªç¹æ®ãªåå¿å®¹åšã
å¿ èŠãšããéåžžã«åŸ®éã®åå¿æ¶²éã§ãããªãããŠ
ãããããã«ãåŸæ¥ã®æ¹æ³ã§ã¯ãå ±åããå®é察
象å€ã®NADãŸãã¯éå åNADè£é µçŽ ãæ¶å»ãã
å Žåãé žãŸãã¯ã¢ã«ã«ãªãæ·»å ãå ç±åŠçåŸãäž
åãããã®ã§ããã®äžåã®ããã®ã¢ã«ã«ãªãŸãã¯
é žã®éãéåžžã«æ£ç¢ºã«çšããŠäžåããããšãèŠ
ããè¯å¥œã«äžåã§ãããPHã«ã°ãã€ããçããå Ž
åãããããµã€ã¯ãªã³ã°çã«å€§ãã圱é¿ããæž¬å®
å€ã®èª€å·®ã®åå ãšãªã€ãŠããããŸãåŸæ¥ã®ATP
ã®æž¬å®æ³ãšããŠã¯ãïŒâãã¹ãã°ãªã»ã¬ã€ãã»ã
ããŒãŒã«ããæ¹æ³ããããœãããŒãŒã«ããæ¹æ³
ïŒMethods of Enzymatic Analysis第ïŒå·»ã第
2097ã2110é ïŒ1974ïŒïŒãç¥ãããŠãããããã
ãã®æ¹æ³ã¯ãATPãšåºè³ªãšã®äœçšã«ãã€ãŠçæ
ãããªã³é žåç©ãããããã²ããŒãŒã«ãŠäœçšãã
ããŠããã®éã«å¿ èŠãšããè£é µçŽ ãäŸãã°éå å
NADïŒïŒ°ïŒã®çæãŸãã¯æžå°éã«ãŠå®éãããã®
ã§ãããããããã®ããããã²ããŒãŒã§ã®å®éã«
åœãããã®åå¿ç³»å ã«ã¯ãäŸãã°NADïŒïŒ°ïŒãšé
å åNADïŒïŒ°ïŒè£é µçŽ ãšãå ±åããå ±åããå®é
察象å€ã®è£é µçŽ ãé žãŸãã¯ã¢ã«ã«ãªã«ãŠç Žå£åŠç
ããããã«äžåããåŸã«ãµã€ã¯ãªã³ã°ãè¡ãããš
ãå¿ é ãšãªããç¹éã§ãã€èªååãåŸãªãæ¹æ³ã§
ããã
ã³ã°æ³ã§ã¯ãäž»åå¿ç³»ã«ã¯NADããã³éå å
NADã®äž¡æ¹ãå ±åããŠããããã®NADããã³é
å åNADã®ããããäžæ¹ãå®éããå Žåã«ã¯ã
ãŸãåå¿ç³»ã«å ±åããå®é察象å€ã®NADãŸãã¯
éå åNADã®ãããããæ¶å»ããã¢ã«ã«ãªãŸã
ã¯é žã®æ·»å ãå ç±åŠçã®æäœãå¿ é ãšããæ¬¡ãã§
PHããµã€ã¯ãªã³ã°åå¿ã«é©ããå€ã«èª¿æŽåŸãµã€ã¯
ãªã³ã°åå¿ãè¡ãªããããã«æç€ºåå¿ããããªã
ãªããã°ãªãããæäœæ§ã«åé¡ãããããã®æç€º
åå¿èªäœã被枬å®ç©è³ªä»¥å€ã®åœ±é¿ãåããå Žåã
å€ãã€ãããããæ²¹äºã®ãããªç¹æ®ãªåå¿å®¹åšã
å¿ èŠãšããéåžžã«åŸ®éã®åå¿æ¶²éã§ãããªãããŠ
ãããããã«ãåŸæ¥ã®æ¹æ³ã§ã¯ãå ±åããå®é察
象å€ã®NADãŸãã¯éå åNADè£é µçŽ ãæ¶å»ãã
å Žåãé žãŸãã¯ã¢ã«ã«ãªãæ·»å ãå ç±åŠçåŸãäž
åãããã®ã§ããã®äžåã®ããã®ã¢ã«ã«ãªãŸãã¯
é žã®éãéåžžã«æ£ç¢ºã«çšããŠäžåããããšãèŠ
ããè¯å¥œã«äžåã§ãããPHã«ã°ãã€ããçããå Ž
åãããããµã€ã¯ãªã³ã°çã«å€§ãã圱é¿ããæž¬å®
å€ã®èª€å·®ã®åå ãšãªã€ãŠããããŸãåŸæ¥ã®ATP
ã®æž¬å®æ³ãšããŠã¯ãïŒâãã¹ãã°ãªã»ã¬ã€ãã»ã
ããŒãŒã«ããæ¹æ³ããããœãããŒãŒã«ããæ¹æ³
ïŒMethods of Enzymatic Analysis第ïŒå·»ã第
2097ã2110é ïŒ1974ïŒïŒãç¥ãããŠãããããã
ãã®æ¹æ³ã¯ãATPãšåºè³ªãšã®äœçšã«ãã€ãŠçæ
ãããªã³é žåç©ãããããã²ããŒãŒã«ãŠäœçšãã
ããŠããã®éã«å¿ èŠãšããè£é µçŽ ãäŸãã°éå å
NADïŒïŒ°ïŒã®çæãŸãã¯æžå°éã«ãŠå®éãããã®
ã§ãããããããã®ããããã²ããŒãŒã§ã®å®éã«
åœãããã®åå¿ç³»å ã«ã¯ãäŸãã°NADïŒïŒ°ïŒãšé
å åNADïŒïŒ°ïŒè£é µçŽ ãšãå ±åããå ±åããå®é
察象å€ã®è£é µçŽ ãé žãŸãã¯ã¢ã«ã«ãªã«ãŠç Žå£åŠç
ããããã«äžåããåŸã«ãµã€ã¯ãªã³ã°ãè¡ãããš
ãå¿ é ãšãªããç¹éã§ãã€èªååãåŸãªãæ¹æ³ã§
ããã
æ¬çºæè
ã¯äžèšã®çš®ã
ã®æ¬ ç¹ã解決ããããã«
è£é µçŽ ãµã€ã¯ãªã³ã°æ³ã«ããå¢å¹ é«æåºŠæž¬å®æ³ã
æ¹è¯ãã¹ãçš®ã ç ç©¶ããçµæããµã€ã¯ãªã³ã°åå¿
èªäœãæç€ºåå¿ãšããããšãããã«ããæç€ºåå¿
ã®ããã®æäœã詊è¬ãå¿ èŠãšããããšã³ãã»ãã€
ã³ãæ³ã ãã§ãªããã¬ã€ãæ³ãå¯èœã§ããç¹åŸŽã
æãããããç¹æ®ãªåå¿å®¹åšãå¿ èŠãšãããäžè¬
ã®ææž©æ§œã§åå¿ãå¯èœã§ããã°ããã§ãªããã©ã€
ã±ãã«ã«æ³ïŒãã€ã«ã æ³ãåºå®åæ³ïŒã«ãå©çšã§
ããéåžžã«ç°¡äŸ¿ãªæ¹æ³ã§ããããšãèŠãåºããã
ããã«æ¬çºæã«ãããŠã¯ãäž»åå¿ç³»ã«ãããŠè£é µ
çŽ ãå¿ èŠãšããªãã®ã§ãäž»åå¿ã«èŠããç©è³ªãæ®
åããŠããŠã被枬å®ç©è³ªã®æž¬å®ã«åœ±é¿ãåãŒããª
ããããäž»åå¿ã®åŸè£é µçŽ ã®äžæŽ»æ§åã®æäœãå¿
èŠãšãããæ£ç¢ºãªæž¬å®ãå¯èœã§ããããšãèŠãåº
ããã
è£é µçŽ ãµã€ã¯ãªã³ã°æ³ã«ããå¢å¹ é«æåºŠæž¬å®æ³ã
æ¹è¯ãã¹ãçš®ã ç ç©¶ããçµæããµã€ã¯ãªã³ã°åå¿
èªäœãæç€ºåå¿ãšããããšãããã«ããæç€ºåå¿
ã®ããã®æäœã詊è¬ãå¿ èŠãšããããšã³ãã»ãã€
ã³ãæ³ã ãã§ãªããã¬ã€ãæ³ãå¯èœã§ããç¹åŸŽã
æãããããç¹æ®ãªåå¿å®¹åšãå¿ èŠãšãããäžè¬
ã®ææž©æ§œã§åå¿ãå¯èœã§ããã°ããã§ãªããã©ã€
ã±ãã«ã«æ³ïŒãã€ã«ã æ³ãåºå®åæ³ïŒã«ãå©çšã§
ããéåžžã«ç°¡äŸ¿ãªæ¹æ³ã§ããããšãèŠãåºããã
ããã«æ¬çºæã«ãããŠã¯ãäž»åå¿ç³»ã«ãããŠè£é µ
çŽ ãå¿ èŠãšããªãã®ã§ãäž»åå¿ã«èŠããç©è³ªãæ®
åããŠããŠã被枬å®ç©è³ªã®æž¬å®ã«åœ±é¿ãåãŒããª
ããããäž»åå¿ã®åŸè£é µçŽ ã®äžæŽ»æ§åã®æäœãå¿
èŠãšãããæ£ç¢ºãªæž¬å®ãå¯èœã§ããããšãèŠãåº
ããã
æ¬çºæã¯äžèšç¥èŠã«åºããã®ã§ãATPããã³
NMNã®ããããäžæåã嫿ããè¢«æ€æ¶²ã«ãäž»
åå¿ç³»ã«ãããŠNMNãŸãã¯ATPãMg2+ã®ååš
äžã«NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒãäœçš
ãããŠNADãšãããªã³é žã«å€æãããäž»åå¿ã®
åŸãçããNADããNADïŒïŒ°ïŒãè£é µçŽ ãšãã
é žåéå åå¿ç³»ãšãéå åNADïŒïŒ°ïŒãè£é µçŽ ãš
ããåå¿ç³»ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯ãªã³ã°
åå¿ããããªããæ¬¡ãã§ãã®ãµã€ã¯ãªã³ã°åå¿ã«
ãããŠçæãŸãã¯æ¶è²»ãããæåãå®éããããš
ãç¹åŸŽãšããè¢«æ€æ¶²äžã®æåã®é«æåºŠæž¬å®æ³ã§ã
ãã
NMNã®ããããäžæåã嫿ããè¢«æ€æ¶²ã«ãäž»
åå¿ç³»ã«ãããŠNMNãŸãã¯ATPãMg2+ã®ååš
äžã«NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒãäœçš
ãããŠNADãšãããªã³é žã«å€æãããäž»åå¿ã®
åŸãçããNADããNADïŒïŒ°ïŒãè£é µçŽ ãšãã
é žåéå åå¿ç³»ãšãéå åNADïŒïŒ°ïŒãè£é µçŽ ãš
ããåå¿ç³»ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯ãªã³ã°
åå¿ããããªããæ¬¡ãã§ãã®ãµã€ã¯ãªã³ã°åå¿ã«
ãããŠçæãŸãã¯æ¶è²»ãããæåãå®éããããš
ãç¹åŸŽãšããè¢«æ€æ¶²äžã®æåã®é«æåºŠæž¬å®æ³ã§ã
ãã
æ¬çºæã«ãããåå¿ç³»ã¯ä»¥äžã®ããã«èª¬æãã
ãã
ãã
äž»åå¿ç³»
ATPïŒNMN
Mg2
ââââ
ããããªã³é
žïŒNAD
NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒ
è£é
µçŽ ãµã€ã¯ãªã³ã°åå¿ç³»ïŒ
NADïŒïŒ°ïŒãè£é
µçŽ ãšããé
žåéå
åå¿
ç³»ïŒ NADïŒS1E1 ââââââââ ãéå åNADïŒP1 éå åNADïŒïŒ°ïŒãè£é µçŽ ãšããåå¿ç³»ïŒ éå åNADïŒS2E2 âââ NADïŒP2 E1ïŒNADããã³S1ãåºè³ªãšããŠæ¶è²»ããé
å åNADããã³P1ãçæããåå¿ãè§Šåª
ããããããã²ããŒãŒã
ç³»ïŒ NADïŒS1E1 ââââââââ ãéå åNADïŒP1 éå åNADïŒïŒ°ïŒãè£é µçŽ ãšããåå¿ç³»ïŒ éå åNADïŒS2E2 âââ NADïŒP2 E1ïŒNADããã³S1ãåºè³ªãšããŠæ¶è²»ããé
å åNADããã³P1ãçæããåå¿ãè§Šåª
ããããããã²ããŒãŒã
E2ïŒéå
åNADããã³S2ãæ¶è²»ããŠãNAD
ããã³P2ãçæããåå¿ãè§Šåªããäœçš
ç©è³ªã
ããã³P2ãçæããåå¿ãè§Šåªããäœçš
ç©è³ªã
S1ïŒE1ã®åºè³ªã
S2ïŒE2ã®åºè³ªã
P1ïŒS1ã®é
žåçæç©ã
P2ïŒS2ã®éå
çæç©ã
ãããå³ç€ºãããšä»¥äžã®ããã«ãªãã
æ¬çºæã®è¢«æ€æ¶²ãšããŠã¯ãå°ãªããšãATPãŸ
ãã¯NMNã®ããããäžæåã嫿ãããã®ã§ã
ãã°ãããATPãŸãã¯NMNãäºã嫿ããŠãª
ãè¢«æ€æ¶²ããATPãŸãã¯NMNãéé¢çæãã
ããŠãªãATPãŸãã¯NMNå«æè¢«æ€æ¶²ãæãã
ãããATPãéé¢çæããããŠãªãATPå«æè¢«
æ€æ¶²ãšããŠã¯ãéåžžãããŒãŒãADPããã³ãã
ãŒãŒåºè³ªçšãªã³ååç©ã«ããé žçŽ åå¿ã«ãŠãã®
ADPããªã³é žåãããŠATPãéé¢ãçæããã
ãé µçŽ åå¿ç³»ã®ãã®ãæãããããããã«è©³ãã
ã¯ãäžèšã®é µçŽ åå¿ç³»ãäŸç€ºããããããããã¯
äŸç€ºã§ãã€ãŠäœããæ¬çºæã®å¯Ÿè±¡ãéå®ãããã®
ã§ã¯ãªãã
ãã¯NMNã®ããããäžæåã嫿ãããã®ã§ã
ãã°ãããATPãŸãã¯NMNãäºã嫿ããŠãª
ãè¢«æ€æ¶²ããATPãŸãã¯NMNãéé¢çæãã
ããŠãªãATPãŸãã¯NMNå«æè¢«æ€æ¶²ãæãã
ãããATPãéé¢çæããããŠãªãATPå«æè¢«
æ€æ¶²ãšããŠã¯ãéåžžãããŒãŒãADPããã³ãã
ãŒãŒåºè³ªçšãªã³ååç©ã«ããé žçŽ åå¿ã«ãŠãã®
ADPããªã³é žåãããŠATPãéé¢ãçæããã
ãé µçŽ åå¿ç³»ã®ãã®ãæãããããããã«è©³ãã
ã¯ãäžèšã®é µçŽ åå¿ç³»ãäŸç€ºããããããããã¯
äŸç€ºã§ãã€ãŠäœããæ¬çºæã®å¯Ÿè±¡ãéå®ãããã®
ã§ã¯ãªãã
ã¯ã¬ã¢ãã³ãããŒãŒïŒEC.2.7.3.2ïŒãADPã
ãã³ã¯ã¬ã¢ãã³ãã¹ããšãŒãã®é µçŽ åå¿ç³»ã§ã
ã¯ã¬ã¢ãã³ãããŒãŒæŽ»æ§æž¬å®ãADPã®å®éã
ã¯ã¬ã¢ãã³ãã¹ããšãŒãã®å®éã®ããããäžæ
åã®æž¬å®ã®ããã«çšããããã
ãã³ã¯ã¬ã¢ãã³ãã¹ããšãŒãã®é µçŽ åå¿ç³»ã§ã
ã¯ã¬ã¢ãã³ãããŒãŒæŽ»æ§æž¬å®ãADPã®å®éã
ã¯ã¬ã¢ãã³ãã¹ããšãŒãã®å®éã®ããããäžæ
åã®æž¬å®ã®ããã«çšããããã
ã¯ã¬ã¢ãã³ãã¹ããšãŒãïŒADP
éå
æžïŒMg2+
âââââââââââ
ã¯ã¬ã¢ãã³ãããŒãŒã¯ã¬ã¢ãã³ïŒATP
ïŒéå
å€ïŒÎ²âã¡ã«ã«ãããšã¿ããŒã«ãéå
å
ã°ã«ã¿ããªã³ãã·ã¹ãã€ã³ãâã¢ã»ãã«ã·ã¹
ãã€ã³ããžããªã¹ã¬ã€ãŒããŒã«ãªã©ïŒ ãã«ããŒããããŒãŒïŒECã2.7.1.40ïŒãADP
ããã³ãã¹ããšããŒã«ãã«ãã³é žã®é žçŽ åå¿ç³»
ã§ããã«ããŒããããŒãŒæŽ»æ§æž¬å®ãADPã®å®
éããã¹ããšããŒã«ãã«ãã³é žã®ããããäžæ
åã®æž¬å®ã«çšããããã
ã°ã«ã¿ããªã³ãã·ã¹ãã€ã³ãâã¢ã»ãã«ã·ã¹
ãã€ã³ããžããªã¹ã¬ã€ãŒããŒã«ãªã©ïŒ ãã«ããŒããããŒãŒïŒECã2.7.1.40ïŒãADP
ããã³ãã¹ããšããŒã«ãã«ãã³é žã®é žçŽ åå¿ç³»
ã§ããã«ããŒããããŒãŒæŽ»æ§æž¬å®ãADPã®å®
éããã¹ããšããŒã«ãã«ãã³é žã®ããããäžæ
åã®æž¬å®ã«çšããããã
ãã¹ããšããŒã«ãã«ãã³é
žïŒADP
Mg2+ãŸãã¯Mn2+ãšK+ïŒNH4 +ãŸãã¯Rb+
âââââââââââââââââââââ
ãã«ããŒããããŒãŒ
ãã«ãã³é
žïŒATP
ã¢ã»ããŒããããŒãŒïŒEC.2.7.2.1ïŒãADPã
ãã³ã¢ã»ãã«ãã¹ããšãŒãã®é µçŽ åå¿ç³»ã§ãã
ã®ããããäžæåã®æž¬å®ã«çšããããã
ãã³ã¢ã»ãã«ãã¹ããšãŒãã®é µçŽ åå¿ç³»ã§ãã
ã®ããããäžæåã®æž¬å®ã«çšããããã
ã¢ã»ãã«ãã¹ããšãŒãïŒADP
Mg2+ãŸãã¯Mn2+
âââââââââââ
ã¢ã»ããŒããããŒãŒé
¢é
žïŒATP
以äžãåé
µçŽ åå¿ç³»ãç°¡ç¥ããŠæããã
ã«ã«ãã¢ã€ã«ãã¹ããšãŒãïŒADP
Mg2+
ââââââââââââââââââââââââ
ã«ã«ãã¡ã€ããããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒ
NH3ïŒCO2ïŒATP
ïŒâãã¹ãââã¢ã¹ãã«ãã€ãïŒADP
Mg2+
Mg2+
âââââââââââââââââââââââââ
ââ ã¢ã¹ãã«ãã€ããããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ âã¢ã¹ãã«ãã€ãïŒATP ïŒïŒïŒâãžãã¹ãââã°ãªã»ã¬ã€ãïŒ
ADP Mg2+ãŸãã¯Mn2+ Mg2+ãŸãã¯Mn2+ âââââââââââââââââââââââââ
âââââ ãã¹ãã°ãªã»ã¬ã€ããããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ ïŒâãã¹ãââã°ãªã»ã¬ã€ãïŒATP ã¢ã«ã®ãã³ãã¹ããšãŒãïŒADP Mg2+ãŸãã¯Mn2+ ââââââââââââââââââââââ ã¢ã«ã®ãã³ãããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ âã¢ã«ã®ãã³ïŒATP ADPïŒADP Mg2+ ââââââââââââââââââââââ ããªãããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ AMPïŒATP XDPïŒADP Mg2+ïŒMn2+ãŸãã¯Ca2+ Mg2+ïŒMn2+ãŸãã¯Ca2+ âââââââââââââââââââââââââ
âââââââââââââââ Mg2+ïŒMn2+ãŸãã¯Ca2+ âââââââââââââââââââââââââ
âââââââââââââââ ãã¯ã¬ãªãµã€ãã¢ããã¹ããšãŒããããŒãŒïŒECïŒïŒïŒïŒ
ïŒïŒïŒïŒïŒ XMPïŒATP ïŒãã ãXDPïŒãã¯ã¬ãªãµã€ããžãã¹ããšãŒ
ãïŒïŒµããããŸãã¯ïŒ¡ã瀺ãïŒ ãããã®é µçŽ åå¿ç³»ã«ããããããŒãŒãšããŠ
ã¯ãäŸãã°åèšã®ã¯ã¬ã¢ãã³ãã¹ããšãŒãããã¹
ããšããŒã«ãã«ãã³é žãã¢ã»ãã«ãã¹ããšãŒãã
ã«ã«ãã¢ã€ã«ãã¹ããšãŒããïŒâãã¹ãââã¢
ã¹ãã«ãã€ããïŒïŒïŒâãžãã¹ãââã°ãªã»ã¬
ã€ããã¢ã«ã®ãã³ãã¹ããšãŒããADPãXDPãª
ã©ã®ãããŒãŒåºè³ªçšãªã³ååç©ã®ãªã³é žåºã
ADPã«è»¢äœããããŠATPãçæéé¢ããäœçšã
æããé µçŽ ã§ããã°ãããäŸãã°ã¯ã¬ã¢ãã³ãã¹
ããšãŒãããããŒãŒåºè³ªçšãªã³ååç©ãšããã¯ã¬
ã¢ãã³ãããŒãŒããã¹ããšããŒã«ãã«ãã³é žãã
ããŒãŒåºè³ªçšãªã³ååç©ãšãããã«ããŒããªãã·
ããŒãŒãã¢ã»ãã«ãã¹ããšãŒãããããŒãŒåºè³ªçš
ãªã³ååç©ãšããã¢ã»ããŒããããŒãŒããã®ä»ã®
ãã¹ãã°ãªã»ã¬ã€ããããŒãŒãã¢ã«ã®ãã³ãããŒ
ãŒãããªãããŒãŒããã¯ã¬ãªãµã€ãã¢ããã¹ããš
ãŒããããŒãŒãªã©ãæãããããNMNãéé¢ç
æããããŠãªãNMNå«æè¢«æ€æ¶²ãšããŠã¯ãäžèš
ã®é µçŽ åå¿ç³»ãäŸç€ºãããããããã¯äŸç€ºã§ãã€
ãŠæ¬çºæã®å¯Ÿè±¡ãäœãéå®ãããã®ã§ã¯ãªãã
ââ ã¢ã¹ãã«ãã€ããããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ âã¢ã¹ãã«ãã€ãïŒATP ïŒïŒïŒâãžãã¹ãââã°ãªã»ã¬ã€ãïŒ
ADP Mg2+ãŸãã¯Mn2+ Mg2+ãŸãã¯Mn2+ âââââââââââââââââââââââââ
âââââ ãã¹ãã°ãªã»ã¬ã€ããããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ ïŒâãã¹ãââã°ãªã»ã¬ã€ãïŒATP ã¢ã«ã®ãã³ãã¹ããšãŒãïŒADP Mg2+ãŸãã¯Mn2+ ââââââââââââââââââââââ ã¢ã«ã®ãã³ãããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ âã¢ã«ã®ãã³ïŒATP ADPïŒADP Mg2+ ââââââââââââââââââââââ ããªãããŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒïŒïŒ AMPïŒATP XDPïŒADP Mg2+ïŒMn2+ãŸãã¯Ca2+ Mg2+ïŒMn2+ãŸãã¯Ca2+ âââââââââââââââââââââââââ
âââââââââââââââ Mg2+ïŒMn2+ãŸãã¯Ca2+ âââââââââââââââââââââââââ
âââââââââââââââ ãã¯ã¬ãªãµã€ãã¢ããã¹ããšãŒããããŒãŒïŒECïŒïŒïŒïŒ
ïŒïŒïŒïŒïŒ XMPïŒATP ïŒãã ãXDPïŒãã¯ã¬ãªãµã€ããžãã¹ããšãŒ
ãïŒïŒµããããŸãã¯ïŒ¡ã瀺ãïŒ ãããã®é µçŽ åå¿ç³»ã«ããããããŒãŒãšããŠ
ã¯ãäŸãã°åèšã®ã¯ã¬ã¢ãã³ãã¹ããšãŒãããã¹
ããšããŒã«ãã«ãã³é žãã¢ã»ãã«ãã¹ããšãŒãã
ã«ã«ãã¢ã€ã«ãã¹ããšãŒããïŒâãã¹ãââã¢
ã¹ãã«ãã€ããïŒïŒïŒâãžãã¹ãââã°ãªã»ã¬
ã€ããã¢ã«ã®ãã³ãã¹ããšãŒããADPãXDPãª
ã©ã®ãããŒãŒåºè³ªçšãªã³ååç©ã®ãªã³é žåºã
ADPã«è»¢äœããããŠATPãçæéé¢ããäœçšã
æããé µçŽ ã§ããã°ãããäŸãã°ã¯ã¬ã¢ãã³ãã¹
ããšãŒãããããŒãŒåºè³ªçšãªã³ååç©ãšããã¯ã¬
ã¢ãã³ãããŒãŒããã¹ããšããŒã«ãã«ãã³é žãã
ããŒãŒåºè³ªçšãªã³ååç©ãšãããã«ããŒããªãã·
ããŒãŒãã¢ã»ãã«ãã¹ããšãŒãããããŒãŒåºè³ªçš
ãªã³ååç©ãšããã¢ã»ããŒããããŒãŒããã®ä»ã®
ãã¹ãã°ãªã»ã¬ã€ããããŒãŒãã¢ã«ã®ãã³ãããŒ
ãŒãããªãããŒãŒããã¯ã¬ãªãµã€ãã¢ããã¹ããš
ãŒããããŒãŒãªã©ãæãããããNMNãéé¢ç
æããããŠãªãNMNå«æè¢«æ€æ¶²ãšããŠã¯ãäžèš
ã®é µçŽ åå¿ç³»ãäŸç€ºãããããããã¯äŸç€ºã§ãã€
ãŠæ¬çºæã®å¯Ÿè±¡ãäœãéå®ãããã®ã§ã¯ãªãã
ãã³ãã³ã¢ããïŒïŒâãã¹ãâαââãª
ãã· ã«ãããã¹ããšãŒã âââââââââââââââââââââââââ
ââââ âââââââââââââââââââââââââ
ââââ NMNãããã¹ããªãªã©ãŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒ12ïŒNMNïŒ
ãããªã³é ž ãããã®é µçŽ åå¿ç³»ã«ãããATPãŸãNMN
ã®å®éç®çã¯ãé µçŽ åå¿ç³»ã«ãããé µçŽ ã®æŽ»æ§æž¬
å®ãçšããããä»ã®æåã®å®éã®ããããäžæå
ã®æž¬å®ã®ããã«è¡ãªããããã®ã§ããŸãé µçŽ åå¿
ç³»ã®æž¬å®æå以å€ã®æåã¯è©Šè¬ãšããŠäžå®éçšã
ãã°ããããã®éçšããããè¢«æ€æ¶²ã詊è¬ã®é
ã¯ã枬å®ãã¹ãç®çãéžæããæ¡ä»¶ã«ãã€ãŠé©å®
èšèšå€æŽããã°ãããç¹ã«éå®ããããã®ã§ã¯ãª
ãããŸããã®é µçŽ åå¿ã§ã¯éåžž37âã§äžåé以äž
è¡ãªãã°ããããã®ããã«æ¬çºæã®è¢«æ€æ¶²ãšããŠ
ã¯ATPãNMNã®ããããäžæåã嫿ããã
ã®ã察象ãšããŠæãããããã®ã§ããã
ãã· ã«ãããã¹ããšãŒã âââââââââââââââââââââââââ
ââââ âââââââââââââââââââââââââ
ââââ NMNãããã¹ããªãªã©ãŒãŒïŒECïŒïŒïŒïŒïŒïŒïŒ12ïŒNMNïŒ
ãããªã³é ž ãããã®é µçŽ åå¿ç³»ã«ãããATPãŸãNMN
ã®å®éç®çã¯ãé µçŽ åå¿ç³»ã«ãããé µçŽ ã®æŽ»æ§æž¬
å®ãçšããããä»ã®æåã®å®éã®ããããäžæå
ã®æž¬å®ã®ããã«è¡ãªããããã®ã§ããŸãé µçŽ åå¿
ç³»ã®æž¬å®æå以å€ã®æåã¯è©Šè¬ãšããŠäžå®éçšã
ãã°ããããã®éçšããããè¢«æ€æ¶²ã詊è¬ã®é
ã¯ã枬å®ãã¹ãç®çãéžæããæ¡ä»¶ã«ãã€ãŠé©å®
èšèšå€æŽããã°ãããç¹ã«éå®ããããã®ã§ã¯ãª
ãããŸããã®é µçŽ åå¿ã§ã¯éåžž37âã§äžåé以äž
è¡ãªãã°ããããã®ããã«æ¬çºæã®è¢«æ€æ¶²ãšããŠ
ã¯ATPãNMNã®ããããäžæåã嫿ããã
ã®ã察象ãšããŠæãããããã®ã§ããã
æ¬çºæã«çšããããNMNã¢ãããªã«ãã©ã³ã¹
ããšã©ãŒãŒã¯ãATPããã³NMNãåºè³ªãšãã
Mg2+ã€ãªã³ã®ååšäžNADããã³ãããªã³é žãç
æããåå¿ãè§Šåªããé µçŽ ïŒEC.2.7.7.1ïŒã§ããã
åç©ãŸãã¯åŸ®çç©ã®ãããã«ç±æ¥ãããã®ã§ãã
ããäŸãã°ãã¿èèãé µæ¯ç±æ¥ã®ãã®ãæããã
ãïŒM.R.AtkinsonãJ.F.JacksonãR.K.
MortonãBiochem.J.ã80ã318ïŒ1981ïŒïŒããã®
NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒã®äœçšã«ã
ããè¢«æ€æ¶²äžã®ATPã¯ãçšããNMNãšãšãã«
NADããã³ãããªã³é žãçæãããŸãè¢«æ€æ¶²äž
ã®NMNã¯çšããATPãšãšãã«NADããã³ãã
ãªã³é žãçæããäž»åå¿ãè¡ãããããã
ããšã©ãŒãŒã¯ãATPããã³NMNãåºè³ªãšãã
Mg2+ã€ãªã³ã®ååšäžNADããã³ãããªã³é žãç
æããåå¿ãè§Šåªããé µçŽ ïŒEC.2.7.7.1ïŒã§ããã
åç©ãŸãã¯åŸ®çç©ã®ãããã«ç±æ¥ãããã®ã§ãã
ããäŸãã°ãã¿èèãé µæ¯ç±æ¥ã®ãã®ãæããã
ãïŒM.R.AtkinsonãJ.F.JacksonãR.K.
MortonãBiochem.J.ã80ã318ïŒ1981ïŒïŒããã®
NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒã®äœçšã«ã
ããè¢«æ€æ¶²äžã®ATPã¯ãçšããNMNãšãšãã«
NADããã³ãããªã³é žãçæãããŸãè¢«æ€æ¶²äž
ã®NMNã¯çšããATPãšãšãã«NADããã³ãã
ãªã³é žãçæããäž»åå¿ãè¡ãããããã
ãã®äž»åå¿ã«ã€ããŠããã®åå¿æ¶²éã¯ãéåžžïŒ
ãã¹ãåœã10ÎŒããïŒmlã®ç¯å²ã®å®¹éã§åå¿ã
åŸããNMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒã¯ã
åå¿æéãŸãã¯åŸ æéã«ããç°ãªãããéåžžïŒã
ã¹ãåœã0.5ã100åäœã奜ãŸããã¯10åäœä»¥äžã§
äœçšãåŸãããŸãçšããããNMNãŸãã¯ATP
ã¯ãå°ãªããšãè¢«æ€æ¶²äžã®ATPãŸãã¯NMNã®
é以äžã«çšããã°ããããŸããã®äž»åå¿ã«ãããŠ
è¢«æ€æ¶²äžã®ATPãŸãã¯NMNã®éã«çžå¿ãã
NADãçæãããã
ãã¹ãåœã10ÎŒããïŒmlã®ç¯å²ã®å®¹éã§åå¿ã
åŸããNMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒã¯ã
åå¿æéãŸãã¯åŸ æéã«ããç°ãªãããéåžžïŒã
ã¹ãåœã0.5ã100åäœã奜ãŸããã¯10åäœä»¥äžã§
äœçšãåŸãããŸãçšããããNMNãŸãã¯ATP
ã¯ãå°ãªããšãè¢«æ€æ¶²äžã®ATPãŸãã¯NMNã®
é以äžã«çšããã°ããããŸããã®äž»åå¿ã«ãããŠ
è¢«æ€æ¶²äžã®ATPãŸãã¯NMNã®éã«çžå¿ãã
NADãçæãããã
次ã«è£é
µçŽ ãµã€ã¯ãªã³ã°ã«ããå¢å¹
åå¿ããã
ã®ã§ããããæ¬çºæã«ãããŠã¯ãäž»åå¿ç³»ã«ãã
ãŠè©Šæäžã®ATPãŸãã¯NMNéã«çžå¿ããŠå€æ
ããçããNADããNADïŒïŒ°ïŒãè£é µçŽ ãšãã
é žåéå åå¿ç³»ãšãéå åNADïŒïŒ°ïŒãè£é µçŽ ãš
ããåå¿ç³»ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯ãªã³ã°
åå¿ããããªããæ¬¡ãã§ãã®ãµã€ã¯ãªã³ã°åå¿ã«
ãããŠçæãŸãã¯æ¶è²»ãããæåãå®éããã
ã®ã§ããããæ¬çºæã«ãããŠã¯ãäž»åå¿ç³»ã«ãã
ãŠè©Šæäžã®ATPãŸãã¯NMNéã«çžå¿ããŠå€æ
ããçããNADããNADïŒïŒ°ïŒãè£é µçŽ ãšãã
é žåéå åå¿ç³»ãšãéå åNADïŒïŒ°ïŒãè£é µçŽ ãš
ããåå¿ç³»ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯ãªã³ã°
åå¿ããããªããæ¬¡ãã§ãã®ãµã€ã¯ãªã³ã°åå¿ã«
ãããŠçæãŸãã¯æ¶è²»ãããæåãå®éããã
NADïŒïŒ°ïŒãšã¯ãNADã®ã¿ããŸãã¯NADãš
NADPã®äž¡æ¹ãšã®ããããã®æå³ã瀺ãããã®
ã§ãNADïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»ãš
ãšããŠã¯äŸãã°ãNADãæ¶è²»ããŠéå åNADã
çæããåå¿ã圢æããããããã²ããŒãŒïŒE1ïŒ
ããã³ãã®åºè³ªïŒS1ïŒã«ããåå¿ç³»ããNADãš
NADPã®äž¡æ¹ãè£é µçŽ ãšããããããã²ããŒãŒ
ïŒE1ïŒããã³ãã®åºè³ªïŒS1ïŒã«ããåå¿ç³»ãçšã
ãããšãã§ãããäžèšã®ããããã²ããŒãŒã¯ãç¹
ã«éå®ãããããšãªããå°ãªããšãNADãè£é µ
çŽ ãšããŠæ¶è²»ãããã®ã§ããã°ããããã€éå°é
çšããç¹å®ã®åºè³ªã«äœçšããŠéå åNADãçæ
ããããããã²ããŒãŒã§ããã°ãããªãèµ·æºã®é µ
çŽ ã§ãã€ãŠãããããããã®é µçŽ ããã³ãã®åºè³ª
ã®äŸãšããŠã¯ãé µçŽ ãã³ãããã¯ãã«èšèŒãããŠ
ããããäŸãšããŠãã©ã¯ããŒãããããã²ããŒãŒ
ïŒEC.1.1.1.27ïŒããã³ïŒ¬âã©ã¯ããŒããã¢ã«ã³ãŒ
ã«ããããã²ããŒãŒïŒEC.1.1.1.1.ïŒããã³ãšã¿ã
ãŒã«ãã°ãªã»ããŒã«ããããã²ããŒãŒ
ïŒEC.1.1.1.6ïŒããã³ã°ãªã»ããŒã«ãã°ãªã»ããŒã«
âïŒâãã¹ããšãŒãããããã²ããŒãŒ
ïŒEC.1.1.1.8ïŒããã³ã°ãªã»ããŒã«âïŒâãã¹ããš
ãŒããã°ã«ã³ãŒã¹ããããã²ããŒãŒ
ïŒEC.1.1.1.47ïŒããã³ã°ã«ã³ãŒã¹ããã¬ãŒããã
ããã²ããŒãŒïŒEC.1.1.1.37ïŒããã³ïŒ¬âãã¬ãŒ
ããã°ã«ã¿ã¡ã€ãããããã²ããŒãŒïŒEC.1.4.1.2ïŒ
ããã³ïŒ¬âã°ã«ã¿ã¡ã€ããïŒâαâããããã·ã¹
ããã€ãããããã²ããŒãŒïŒEC.1.1.1.50ïŒããã³
ïŒâαâããããã·ã¹ããã€ããªã©ãæããã
ãããããã®é µçŽ ãçšããé žåéå åå¿ç³»ã«ãã
ãé µçŽ ïŒE1ïŒãåºè³ªïŒS1ïŒãé µçŽ åå¿ãçæç©
ïŒP1ïŒã¯ä»¥äžã«ç€ºãéãã§ããã
NADPã®äž¡æ¹ãšã®ããããã®æå³ã瀺ãããã®
ã§ãNADïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»ãš
ãšããŠã¯äŸãã°ãNADãæ¶è²»ããŠéå åNADã
çæããåå¿ã圢æããããããã²ããŒãŒïŒE1ïŒ
ããã³ãã®åºè³ªïŒS1ïŒã«ããåå¿ç³»ããNADãš
NADPã®äž¡æ¹ãè£é µçŽ ãšããããããã²ããŒãŒ
ïŒE1ïŒããã³ãã®åºè³ªïŒS1ïŒã«ããåå¿ç³»ãçšã
ãããšãã§ãããäžèšã®ããããã²ããŒãŒã¯ãç¹
ã«éå®ãããããšãªããå°ãªããšãNADãè£é µ
çŽ ãšããŠæ¶è²»ãããã®ã§ããã°ããããã€éå°é
çšããç¹å®ã®åºè³ªã«äœçšããŠéå åNADãçæ
ããããããã²ããŒãŒã§ããã°ãããªãèµ·æºã®é µ
çŽ ã§ãã€ãŠãããããããã®é µçŽ ããã³ãã®åºè³ª
ã®äŸãšããŠã¯ãé µçŽ ãã³ãããã¯ãã«èšèŒãããŠ
ããããäŸãšããŠãã©ã¯ããŒãããããã²ããŒãŒ
ïŒEC.1.1.1.27ïŒããã³ïŒ¬âã©ã¯ããŒããã¢ã«ã³ãŒ
ã«ããããã²ããŒãŒïŒEC.1.1.1.1.ïŒããã³ãšã¿ã
ãŒã«ãã°ãªã»ããŒã«ããããã²ããŒãŒ
ïŒEC.1.1.1.6ïŒããã³ã°ãªã»ããŒã«ãã°ãªã»ããŒã«
âïŒâãã¹ããšãŒãããããã²ããŒãŒ
ïŒEC.1.1.1.8ïŒããã³ã°ãªã»ããŒã«âïŒâãã¹ããš
ãŒããã°ã«ã³ãŒã¹ããããã²ããŒãŒ
ïŒEC.1.1.1.47ïŒããã³ã°ã«ã³ãŒã¹ããã¬ãŒããã
ããã²ããŒãŒïŒEC.1.1.1.37ïŒããã³ïŒ¬âãã¬ãŒ
ããã°ã«ã¿ã¡ã€ãããããã²ããŒãŒïŒEC.1.4.1.2ïŒ
ããã³ïŒ¬âã°ã«ã¿ã¡ã€ããïŒâαâããããã·ã¹
ããã€ãããããã²ããŒãŒïŒEC.1.1.1.50ïŒããã³
ïŒâαâããããã·ã¹ããã€ããªã©ãæããã
ãããããã®é µçŽ ãçšããé žåéå åå¿ç³»ã«ãã
ãé µçŽ ïŒE1ïŒãåºè³ªïŒS1ïŒãé µçŽ åå¿ãçæç©
ïŒP1ïŒã¯ä»¥äžã«ç€ºãéãã§ããã
âã©ã¯ããŒãïŒS1ïŒïŒNAD
ïŒE1ïŒ
âââââââââââââââ
ã©ã¯ããŒãããããã²ããŒãŒ
ãã«ãã³é
žïŒP1ïŒïŒéå
åNAD
ãšã¿ããŒã«ïŒS1ïŒïŒNAD
ïŒE1ïŒ
âââââââââââââââ
ã¢ã«ã³ãŒã«ããããã²ããŒãŒ
ã¢ã»ãã¢ã«ãããïŒP1ïŒïŒéå
åNAD
ã°ãªã»ããŒã«ïŒS1ïŒïŒNAD
ïŒE1ïŒ
ââââââââââââââââ
ã°ãªã»ããŒã«ããããã²ããŒãŒ
ãžããããã·ã¢ã»ãã³ïŒP1ïŒïŒéå
åNAD
ã°ãªã»ããŒã«âïŒâãã¹ããšãŒã
ïŒS1ïŒïŒNAD
ïŒE1ïŒ
ââââââââââ
ã°ãªã»ããŒã«âïŒâãã¹ããšãŒã
ããããã²ããŒãŒ
ãžããããã·ã¢ã»ãã³ãã¹ããšãŒãïŒP1ïŒ
ïŒéå
åNAD
ã°ã«ã³ãŒã¹ïŒS1ïŒïŒNADïŒïŒ°ïŒ
ïŒE1ïŒ
âââââââââââââââ
ã°ã«ã³ãŒã¹ããããã²ããŒãŒ
ã°ã«ã³ãâÎŽâã©ã¯ãã³ïŒP1ïŒïŒéå
åNAD
âãã¬ãŒãïŒS1ïŒïŒNAD
ïŒE1ïŒ
ââââââââââââââ
ãã¬ãŒãããããã²ããŒãŒ
ãªã®ã¶ãé
¢é
žïŒP1ïŒïŒéå
åNAD
âã°ã«ã¿ã¡ã€ãïŒS1ïŒïŒH2OïŒNAD
ïŒE1ïŒ
ââââââââââââââââ
ã°ã«ã¿ã¡ã€ãããããã²ããŒãŒ
ïŒâãªããœã°ã«ã¿ã¬ã€ãïŒP1ïŒïŒéå
åNAD
ïŒâαâããããã·ã¹ããã€ãïŒS1ïŒïŒNAD
ïŒE1ïŒ
âââââââââââââââ
ïŒâαâããããã·ã¹ã©ãã€ã
ããããã²ããŒãŒ
ïŒâã±ãã¹ããã€ãïŒP1ïŒïŒéå
åNAD
ãããã®é
žåéå
åå¿ã«äœ¿çšããé
µçŽ éã¯ãé
µ
çŽ å䟡ãåºè³ªã®çš®é¡ããã³è£é µçŽ ãµã€ã¯ãªã³ã°ç
ã«ãã€ãŠç°ãªããåºè³ªéã¯ãïŒãµã€ã¯ã«æ¯ã«ïŒã¢
ã«æ¯ã®åºè³ªãæ¶è²»ããŠãªããã®ã§ããµã€ã¯ãªã³ã°
ããè£é µçŽ ããæ¯èŒã«ãªããªãçšã¯ããã«å€ãã¢
ã«éã䜿çšããããã®ã§ãéåžžåäœæéåœãã®ãµ
ã€ã¯ã«æ°ããã³åå¿æéã«åºããŠæ±ºå®ããã°ã
ãããŸããã®é žåéå é µçŽ ã®åå¿é床ãæå€§ã瀺
ããããªæ¿åºŠä»¥äžã§ããã°ãããéåžž0.1ïœïŒãª
ãã100ïœïŒã®æ¿åºŠç¯å²ã§ååšãåŸãã
çŽ å䟡ãåºè³ªã®çš®é¡ããã³è£é µçŽ ãµã€ã¯ãªã³ã°ç
ã«ãã€ãŠç°ãªããåºè³ªéã¯ãïŒãµã€ã¯ã«æ¯ã«ïŒã¢
ã«æ¯ã®åºè³ªãæ¶è²»ããŠãªããã®ã§ããµã€ã¯ãªã³ã°
ããè£é µçŽ ããæ¯èŒã«ãªããªãçšã¯ããã«å€ãã¢
ã«éã䜿çšããããã®ã§ãéåžžåäœæéåœãã®ãµ
ã€ã¯ã«æ°ããã³åå¿æéã«åºããŠæ±ºå®ããã°ã
ãããŸããã®é žåéå é µçŽ ã®åå¿é床ãæå€§ã瀺
ããããªæ¿åºŠä»¥äžã§ããã°ãããéåžž0.1ïœïŒãª
ãã100ïœïŒã®æ¿åºŠç¯å²ã§ååšãåŸãã
éå
åNADïŒïŒ°ïŒãè£é
µçŽ ãšããåå¿ç³»ãšããŠ
ã¯ãå°ãªããšãéå åNADãæ¶è²»ããŠNADãç
æããäœçšç©è³ªïŒE2ïŒããã³ãã®åºè³ªïŒS2ïŒã®
åå¿ç³»ãæãããããã®äœçšç©è³ªãšãã®åºè³ªã®å
å¿ç³»ãšããŠã¯ãå°ãªããšãéå åNADãæ¶è²»ã
ãŠNADãçæããé žåéå é µçŽ ããã³ãã®åºè³ª
ã®åå¿ç³»ããé»åäŒéç©è³ªããã³ããã©ãŸãªãŠã
å¡©ã®åå¿ç³»ãªã©ãæããããã
ã¯ãå°ãªããšãéå åNADãæ¶è²»ããŠNADãç
æããäœçšç©è³ªïŒE2ïŒããã³ãã®åºè³ªïŒS2ïŒã®
åå¿ç³»ãæãããããã®äœçšç©è³ªãšãã®åºè³ªã®å
å¿ç³»ãšããŠã¯ãå°ãªããšãéå åNADãæ¶è²»ã
ãŠNADãçæããé žåéå é µçŽ ããã³ãã®åºè³ª
ã®åå¿ç³»ããé»åäŒéç©è³ªããã³ããã©ãŸãªãŠã
å¡©ã®åå¿ç³»ãªã©ãæããããã
éå
åNADãæ¶è²»ããŠNADãçæããé
žåé
å é µçŽ ãšããŠã¯ãå°ãªããšãéå åNADãè£é µ
çŽ ãšããéå°éçšããç¹å®ã®åºè³ªïŒS2ïŒã«äœçšã
ãŠNADããã³S2ã®éå åçæç©ïŒP2ïŒãçæã
ãããããã²ããŒãŒããŸãã¯å°ãªããšãéå å
NADãè£é µçŽ ãšããããã¯ããŒã ããžã¹ã«ãã€
ãååç©ãããã³ããã³ãã®é¡çžäœçãå容äœãš
ããéå åNADïŒïŒ°ïŒïŒå容äœé žåéå é µçŽ ã§ã
ãã°ãã®ãããã§ãè¯ãããã®èµ·æºãéå®ããã
ããšã¯ãªãããããã®é µçŽ ããã³ãã®åºè³ªãŸãã¯
å容äœã®äŸãšããŠã¯ããé µçŽ ãã³ãããã¯ãã«èš
èŒãããŠããããããããã²ããŒãŒããã³ãã®åº
質ã®äŸãšããŠã¯ãã©ã¯ããŒãããããã²ããŒãŒ
ïŒEC.1.1.1.27ïŒããã³ãã«ãã³é žãã¢ã«ã³ãŒã«ã
ãããã²ããŒãŒïŒEC.1.1.1.1ïŒããã³ã¢ã»ãã¢ã«
ããããã°ãªã»ããŒã«ããããã²ããŒãŒ
ïŒEC.1.1.1.6ïŒããã³ãžããããã·ã¢ã»ãã³ãã°ãª
ã»ããŒã«âïŒâãã¹ããšãŒãããããã²ããŒãŒ
ïŒEC.1.1.1.8ïŒããã³ãžããããã·ã¢ã»ãã³ãã¹ã
ãšãŒãïŒããã¬ãŒãããããã²ããŒãŒ
ïŒEC.1.1.1.37ïŒããã³ãªã®ã¶ãé ¢é žãïŒâαâã
ãããã·ã¹ããã€ãããããã²ããŒãŒ
ïŒEC.1.1.1.50ïŒããã³ïŒâã±ãâã¹ããã€ããªã©
ã®äŸãæããããããŸãéå åNADïŒïŒ°ïŒïŒå容
äœé žåéå é µçŽ ãšããŠã¯ãããã¯ããŒã b5ã¬ãã¯
ã¿ãŒãŒïŒEC.1.6.2.2ïŒããžã¢ãã©ãŒãŒ
ïŒEC.1.6.4.3ïŒããã€ãã¬ãŒãã¬ãã¯ã¿ãŒãŒ
ïŒEC.1.6.6.1ïŒãéå åNADPããããã²ããŒãŒ
ïŒEC.1.6.99.1ïŒãéå åNADïŒïŒ°ïŒããããã²ããŒ
ãŒïŒEC.1.6.99.2ïŒãéå åNADããããã²ããŒãŒ
ïŒEC.1.6.99.3ïŒãéå åNADããããã²ããŒãŒ
ïŒããã³ïŒïŒEC.1.6.99.5ïŒãªã©ãæãããããå容
äœãšããŠã¯ãã¡ãã¬ã³ã»ãã«ãŒããã©ãã³é¡ãã
ãã³é¡ãïŒïŒïŒâãžã¯ããããšããŒã«ã€ã³ãããš
ããŒã«ãããšãªã·ã¢ã³åå¡©ãããã©ãŸãªãŠã å¡©ã
ããã¯ããŒã ããªã©ãçšãããåŸãããæŽã«å ·äœ
çã«ã¯ããã©ãã³é¡ãšããŠã¯ãªããã©ãã³ããã©
ãã³ã¢ããã¯ã¬ãªããïŒFMNïŒããã©ãã³ã¢ã
ãã³ãžãã¯ã¬ãªããïŒFADïŒãããã³é¡ãšããŠã¯
ãããããã³é¡ãšããŠãïŒïŒïŒâãããããã³ã
ïŒâã¡ãã«âïŒïŒïŒâãããããã³ããã³ãŸãã
ã³é¡ãšããŠã¯ïŒïŒïŒâãžã¡ããã·âïŒâã¡ãã«â
ïŒïŒïŒâãã³ãŸããã³ãããã©ã¡ãã«ââãã³
ãŸããã³ãâãã³ãŸããã³ããŠãããã³ããã
ã©ãŸãªãŠã å¡©ãšããŠã¯ïŒïŒ3â²âïŒïŒïŒ3â²âãžã¡ã
ãã·âïŒïŒ4â²âãžããšãã¬ã³ïŒãã¹ãïŒâïŒïŒ°â
ãããããšãã«ïŒâïŒâïŒâããšãã«âããã©ãŸ
ãªãŠã ã¯ãã©ã€ããïŒãããããã©ãŸãªãŠã ãã«
ãŒïŒïŒNTBïŒãïŒâïŒïŒ°âãããããšãã«ïŒâïŒâ
ïŒïŒ°âãšãŒãããšãã«ïŒâïŒâããšãã«ããã©ãŸãª
ãŠã ã¯ãã©ã€ãïŒINTïŒãïŒâïŒ4â²ïŒ5â²âãžã¡ãã«
â2â²âãã¢ãŸãªã«ïŒâïŒïŒïŒâãžããšãã«ããã©
ãŸãªãŠã ãããã€ãïŒïŒïŒïŒâMTTïŒãããã¯
ããŒã ãšããŠã¯ããšãªããã¯ããŒã b5ãããã¯ã
ãŒã ãªã©ãæããããã
å é µçŽ ãšããŠã¯ãå°ãªããšãéå åNADãè£é µ
çŽ ãšããéå°éçšããç¹å®ã®åºè³ªïŒS2ïŒã«äœçšã
ãŠNADããã³S2ã®éå åçæç©ïŒP2ïŒãçæã
ãããããã²ããŒãŒããŸãã¯å°ãªããšãéå å
NADãè£é µçŽ ãšããããã¯ããŒã ããžã¹ã«ãã€
ãååç©ãããã³ããã³ãã®é¡çžäœçãå容äœãš
ããéå åNADïŒïŒ°ïŒïŒå容äœé žåéå é µçŽ ã§ã
ãã°ãã®ãããã§ãè¯ãããã®èµ·æºãéå®ããã
ããšã¯ãªãããããã®é µçŽ ããã³ãã®åºè³ªãŸãã¯
å容äœã®äŸãšããŠã¯ããé µçŽ ãã³ãããã¯ãã«èš
èŒãããŠããããããããã²ããŒãŒããã³ãã®åº
質ã®äŸãšããŠã¯ãã©ã¯ããŒãããããã²ããŒãŒ
ïŒEC.1.1.1.27ïŒããã³ãã«ãã³é žãã¢ã«ã³ãŒã«ã
ãããã²ããŒãŒïŒEC.1.1.1.1ïŒããã³ã¢ã»ãã¢ã«
ããããã°ãªã»ããŒã«ããããã²ããŒãŒ
ïŒEC.1.1.1.6ïŒããã³ãžããããã·ã¢ã»ãã³ãã°ãª
ã»ããŒã«âïŒâãã¹ããšãŒãããããã²ããŒãŒ
ïŒEC.1.1.1.8ïŒããã³ãžããããã·ã¢ã»ãã³ãã¹ã
ãšãŒãïŒããã¬ãŒãããããã²ããŒãŒ
ïŒEC.1.1.1.37ïŒããã³ãªã®ã¶ãé ¢é žãïŒâαâã
ãããã·ã¹ããã€ãããããã²ããŒãŒ
ïŒEC.1.1.1.50ïŒããã³ïŒâã±ãâã¹ããã€ããªã©
ã®äŸãæããããããŸãéå åNADïŒïŒ°ïŒïŒå容
äœé žåéå é µçŽ ãšããŠã¯ãããã¯ããŒã b5ã¬ãã¯
ã¿ãŒãŒïŒEC.1.6.2.2ïŒããžã¢ãã©ãŒãŒ
ïŒEC.1.6.4.3ïŒããã€ãã¬ãŒãã¬ãã¯ã¿ãŒãŒ
ïŒEC.1.6.6.1ïŒãéå åNADPããããã²ããŒãŒ
ïŒEC.1.6.99.1ïŒãéå åNADïŒïŒ°ïŒããããã²ããŒ
ãŒïŒEC.1.6.99.2ïŒãéå åNADããããã²ããŒãŒ
ïŒEC.1.6.99.3ïŒãéå åNADããããã²ããŒãŒ
ïŒããã³ïŒïŒEC.1.6.99.5ïŒãªã©ãæãããããå容
äœãšããŠã¯ãã¡ãã¬ã³ã»ãã«ãŒããã©ãã³é¡ãã
ãã³é¡ãïŒïŒïŒâãžã¯ããããšããŒã«ã€ã³ãããš
ããŒã«ãããšãªã·ã¢ã³åå¡©ãããã©ãŸãªãŠã å¡©ã
ããã¯ããŒã ããªã©ãçšãããåŸãããæŽã«å ·äœ
çã«ã¯ããã©ãã³é¡ãšããŠã¯ãªããã©ãã³ããã©
ãã³ã¢ããã¯ã¬ãªããïŒFMNïŒããã©ãã³ã¢ã
ãã³ãžãã¯ã¬ãªããïŒFADïŒãããã³é¡ãšããŠã¯
ãããããã³é¡ãšããŠãïŒïŒïŒâãããããã³ã
ïŒâã¡ãã«âïŒïŒïŒâãããããã³ããã³ãŸãã
ã³é¡ãšããŠã¯ïŒïŒïŒâãžã¡ããã·âïŒâã¡ãã«â
ïŒïŒïŒâãã³ãŸããã³ãããã©ã¡ãã«ââãã³
ãŸããã³ãâãã³ãŸããã³ããŠãããã³ããã
ã©ãŸãªãŠã å¡©ãšããŠã¯ïŒïŒ3â²âïŒïŒïŒ3â²âãžã¡ã
ãã·âïŒïŒ4â²âãžããšãã¬ã³ïŒãã¹ãïŒâïŒïŒ°â
ãããããšãã«ïŒâïŒâïŒâããšãã«âããã©ãŸ
ãªãŠã ã¯ãã©ã€ããïŒãããããã©ãŸãªãŠã ãã«
ãŒïŒïŒNTBïŒãïŒâïŒïŒ°âãããããšãã«ïŒâïŒâ
ïŒïŒ°âãšãŒãããšãã«ïŒâïŒâããšãã«ããã©ãŸãª
ãŠã ã¯ãã©ã€ãïŒINTïŒãïŒâïŒ4â²ïŒ5â²âãžã¡ãã«
â2â²âãã¢ãŸãªã«ïŒâïŒïŒïŒâãžããšãã«ããã©
ãŸãªãŠã ãããã€ãïŒïŒïŒïŒâMTTïŒãããã¯
ããŒã ãšããŠã¯ããšãªããã¯ããŒã b5ãããã¯ã
ãŒã ãªã©ãæããããã
éå
åNADïŒïŒ°ïŒïŒå容äœé
žåéå
é
µçŽ ããã³
å容äœã®çµåãã¯å°ãªããšãéå åNADãè£é µ
çŽ ãšããŠãªãé µçŽ ããã³é»åå容äœãšãªãåŸãã
ã®ã§ããã°ç¹ã«å¶éãããªãã奜ãŸããçµã¿åã
ãšããŠã¯ããžã¢ãã©ãŒãŒïŒEC.1.6.4.3ïŒããã³ã
ãã©ãŸãªãŠã å¡©ãéå åNADPããããã²ããŒ
ãŒïŒEC.1.6.99.1ïŒããã³ã¡ãã¬ã³ã»ãã«ãŒãéå
åNADããããã²ããŒãŒïŒEC.1.6.99.3ïŒããã³
ããã¯ããŒã ãªã©ãæãããããéå åNAD
ïŒïŒ°ïŒïŒå容äœé žåé µçŽ ã¯éå åNADã«ç¹ç°æ§ã®
é«ããã®ã奜ãŸãããäœ¿çšæ¿åºŠã¯éåžž0.05ã
100UïŒmlã®ç¯å²ã§ååšãåŸããããã©ãŸãªãŠã
å¡©ã®äœ¿çšæ¿åºŠã¯ãããã©ãŸãªãŠã å¡©ããã³ç©¶æ¥µç
ã«åœ¢æããããã«ãã¶ã³ã®åæ¹ã®æ°Žæº¶æ§ãããã
éå®ãããããéåžžã¯è©Šè¬ïŒmlåœã1ÎŒïœãã
100ÎŒïœã®æ¿åºŠç¯å²ã§ååšãåŸãã
å容äœã®çµåãã¯å°ãªããšãéå åNADãè£é µ
çŽ ãšããŠãªãé µçŽ ããã³é»åå容äœãšãªãåŸãã
ã®ã§ããã°ç¹ã«å¶éãããªãã奜ãŸããçµã¿åã
ãšããŠã¯ããžã¢ãã©ãŒãŒïŒEC.1.6.4.3ïŒããã³ã
ãã©ãŸãªãŠã å¡©ãéå åNADPããããã²ããŒ
ãŒïŒEC.1.6.99.1ïŒããã³ã¡ãã¬ã³ã»ãã«ãŒãéå
åNADããããã²ããŒãŒïŒEC.1.6.99.3ïŒããã³
ããã¯ããŒã ãªã©ãæãããããéå åNAD
ïŒïŒ°ïŒïŒå容äœé žåé µçŽ ã¯éå åNADã«ç¹ç°æ§ã®
é«ããã®ã奜ãŸãããäœ¿çšæ¿åºŠã¯éåžž0.05ã
100UïŒmlã®ç¯å²ã§ååšãåŸããããã©ãŸãªãŠã
å¡©ã®äœ¿çšæ¿åºŠã¯ãããã©ãŸãªãŠã å¡©ããã³ç©¶æ¥µç
ã«åœ¢æããããã«ãã¶ã³ã®åæ¹ã®æ°Žæº¶æ§ãããã
éå®ãããããéåžžã¯è©Šè¬ïŒmlåœã1ÎŒïœãã
100ÎŒïœã®æ¿åºŠç¯å²ã§ååšãåŸãã
é»åäŒéç©è³ªãšããŠã¯ãéå
åNADãNADã«
é žåããèœåãæãããããè£é µçŽ ãµã€ã¯ãªã³ã°
åå¿ã«æªåœ±é¿ãåãŒããªããããªç©è³ªã§ãããäŸ
ãã°ããšãã·ã³ã¡ã¿ã«ãµã«ããšãŒããã¡ã«ããŒ
ã©ã»ãã«ãŒãããã·ã¢ãã³ãªã©ãæããããã䜿
çšæ¿åºŠã¯ããµã€ã¯ãªã³ã°çã«å¿ããŠèšå®ããã°ã
ãããéåžžåå¿æ¶²ïŒmlåœã5ÎŒïœã0.5mgã®æ¿åºŠç¯
å²ã§ååšãåŸãã
é žåããèœåãæãããããè£é µçŽ ãµã€ã¯ãªã³ã°
åå¿ã«æªåœ±é¿ãåãŒããªããããªç©è³ªã§ãããäŸ
ãã°ããšãã·ã³ã¡ã¿ã«ãµã«ããšãŒããã¡ã«ããŒ
ã©ã»ãã«ãŒãããã·ã¢ãã³ãªã©ãæããããã䜿
çšæ¿åºŠã¯ããµã€ã¯ãªã³ã°çã«å¿ããŠèšå®ããã°ã
ãããéåžžåå¿æ¶²ïŒmlåœã5ÎŒïœã0.5mgã®æ¿åºŠç¯
å²ã§ååšãåŸãã
äžèšã®ãµã€ã¯ãªã³ã°åå¿ã¯ãé垞宀枩ãªãã37
âä»è¿ã®æž©åºŠã奜ãŸããã¯30ã37âã®æž©åºŠã§è¡ã
ãããåå¿æéã¯ãç¹ã«éå®ããããã®ã§ãªãã
éåžžïŒå以äžã奜ãŸããã¯ïŒå以äžè¡ãªãã°ã
ããäºå®ãããæéã®çµãã§åå¿ãè¿ éã«åæ¢ã
ãã«ã¯ãé žãäŸãã°å¡©é žããªã³é žãªã©ãæ·»å ãã
ããšã«ããè¡ãªãããã
âä»è¿ã®æž©åºŠã奜ãŸããã¯30ã37âã®æž©åºŠã§è¡ã
ãããåå¿æéã¯ãç¹ã«éå®ããããã®ã§ãªãã
éåžžïŒå以äžã奜ãŸããã¯ïŒå以äžè¡ãªãã°ã
ããäºå®ãããæéã®çµãã§åå¿ãè¿ éã«åæ¢ã
ãã«ã¯ãé žãäŸãã°å¡©é žããªã³é žãªã©ãæ·»å ãã
ããšã«ããè¡ãªãããã
ãã®ããã«ããŠãµã€ã¯ãªã³ã°åå¿ãè¡ãªã€ã
åŸããã®ãµã€ã¯ãªã³ã°åå¿ã«ãããŠçæãŸãã¯æ¶
è²»ãããéšåãå®éããã®ã§ããããçæããæ
åãšããŠã¯E1ã®åºè³ªïŒS1ïŒã®é žåçæç©ïŒP1ïŒ
ãŸãã¯E2ã®åºè³ªïŒS2ïŒã®éå çæç©ïŒP2ïŒã察
象ãšããã°ãããæ¶è²»ãããæåãšããŠã¯E1ã®
åºè³ªïŒS1ïŒãŸãã¯E2ã®åºè³ªïŒS2ïŒã察象ãšãã
ã°ããããããã®P1ãP2ãS1ãS2ã®ããããäž
æåã®éãå®éããã°ããã簡䟿ã«ã¯ãåºè³ªã®ç¶
æ ã§ã¯ç¡è²ã§ãããçæç©ã®ç¶æ ã«ãŠåè²ãŸãã¯
è¢å ãåããåžå æ³¢é·ãå€åããå Žåã®çæç©ã®
å®éææ®µãçšããããšã§ãããäŸãã°ããã©ãŸãª
ãŠã å¡©ãåºè³ªïŒS2ïŒãšããŠãçæãããã«ãã¶ã³
ãéå çæç©ïŒP2ïŒãšããããŠããã®ãã«ãã¶
ã³ãæ¯è²å®éããŠãªããã®ã§ãããããã«ãã©ã
ã³é¡ãããã³é¡ãåºè³ªïŒS2ïŒãšããŠçšããå Žåã«
ã¯ããããã®åºè³ªïŒïŒ³ïŒã®æ¶è²»éããã®ç¹æã®åž
å æ³¢é·ã«åºããŠåžå 床枬å®ããŠå®éããã°ããã
åŸããã®ãµã€ã¯ãªã³ã°åå¿ã«ãããŠçæãŸãã¯æ¶
è²»ãããéšåãå®éããã®ã§ããããçæããæ
åãšããŠã¯E1ã®åºè³ªïŒS1ïŒã®é žåçæç©ïŒP1ïŒ
ãŸãã¯E2ã®åºè³ªïŒS2ïŒã®éå çæç©ïŒP2ïŒã察
象ãšããã°ãããæ¶è²»ãããæåãšããŠã¯E1ã®
åºè³ªïŒS1ïŒãŸãã¯E2ã®åºè³ªïŒS2ïŒã察象ãšãã
ã°ããããããã®P1ãP2ãS1ãS2ã®ããããäž
æåã®éãå®éããã°ããã簡䟿ã«ã¯ãåºè³ªã®ç¶
æ ã§ã¯ç¡è²ã§ãããçæç©ã®ç¶æ ã«ãŠåè²ãŸãã¯
è¢å ãåããåžå æ³¢é·ãå€åããå Žåã®çæç©ã®
å®éææ®µãçšããããšã§ãããäŸãã°ããã©ãŸãª
ãŠã å¡©ãåºè³ªïŒS2ïŒãšããŠãçæãããã«ãã¶ã³
ãéå çæç©ïŒP2ïŒãšããããŠããã®ãã«ãã¶
ã³ãæ¯è²å®éããŠãªããã®ã§ãããããã«ãã©ã
ã³é¡ãããã³é¡ãåºè³ªïŒS2ïŒãšããŠçšããå Žåã«
ã¯ããããã®åºè³ªïŒïŒ³ïŒã®æ¶è²»éããã®ç¹æã®åž
å æ³¢é·ã«åºããŠåžå 床枬å®ããŠå®éããã°ããã
ããã«äžèšåå¿ã«ãããŠãããã©ãŸãªãŠã å¡©ã
ã圢æããããã«ãã¶ã³ã®æ²æŸ±ã®é²æ¢ããããã
ããã«ç颿޻æ§å€ãæ·»å ããããšã奜ãŸãããç
颿޻æ§å€ãšããŠã¯ãã©ã€ãã³ïŒžâ100ãã¢ãã«ã
ãŒã«SOâ145ãªã©ã®éã€ãªã³ç颿޻æ§å€ãæãã
ãããäœ¿çšæ¿åºŠã¯è©Šè¬ã«å¯Ÿãã0.01ãïŒïŒ ã®æ¿åºŠ
ç¯å²ã§ååšãåŸãããã®ç颿޻æ§å€ã®æ·»å ã«ã
ããæž¬å®å€ã®æåºŠã®äžæãšãã«ãã¶ã³è²çŽ ã®å®å®
åãèšãããšãã§ããã
ã圢æããããã«ãã¶ã³ã®æ²æŸ±ã®é²æ¢ããããã
ããã«ç颿޻æ§å€ãæ·»å ããããšã奜ãŸãããç
颿޻æ§å€ãšããŠã¯ãã©ã€ãã³ïŒžâ100ãã¢ãã«ã
ãŒã«SOâ145ãªã©ã®éã€ãªã³ç颿޻æ§å€ãæãã
ãããäœ¿çšæ¿åºŠã¯è©Šè¬ã«å¯Ÿãã0.01ãïŒïŒ ã®æ¿åºŠ
ç¯å²ã§ååšãåŸãããã®ç颿޻æ§å€ã®æ·»å ã«ã
ããæž¬å®å€ã®æåºŠã®äžæãšãã«ãã¶ã³è²çŽ ã®å®å®
åãèšãããšãã§ããã
çãããã«ãã¶ã³è²çŽ ã®æ¯è²å®éã¯ãã«ãã¶ã³
ã®ç¹ç°çåžå æ³¢é·ã«ãŠåžå 床ïŒODïŒã枬å®ãã
ã°ãããäŸãã°ã500ã550nïœã®æ³¢é·ã«ããåžå
åºŠãæž¬å®ããŠã被枬å®ç©è³ªã®å®éãè¡ãããšãã§
ããã
ã®ç¹ç°çåžå æ³¢é·ã«ãŠåžå 床ïŒODïŒã枬å®ãã
ã°ãããäŸãã°ã500ã550nïœã®æ³¢é·ã«ããåžå
åºŠãæž¬å®ããŠã被枬å®ç©è³ªã®å®éãè¡ãããšãã§
ããã
æ¬çºæã«ããã°ããšã³ãã»ãã€ã³ãæ³ã ãã§ãª
ããã¬ã€ãæ³ããã©ã€ã±ãã«ã«æ³ïŒãã€ã«ã æ³ã
åºå®åæ³ïŒãå¯èœã§ããã
ããã¬ã€ãæ³ããã©ã€ã±ãã«ã«æ³ïŒãã€ã«ã æ³ã
åºå®åæ³ïŒãå¯èœã§ããã
æ¬çºæã¯ATPã®é«æåºŠæž¬å®æ³ãšããŠæçšã§ã
ãã詊æäžã«éé¢ããŠããATPã®å®éãããã
ãŒãŒãADPããã³ãããŒãŒåºè³ªçšãªã³ååç©ã«
ããé µçŽ åå¿ã«ãŠADPããªã³é žåãããŠçæã
éé¢ãããATPã®å®éãNMNã®å®éãNMNã
ããã¹ããªã©ãŒãŒããã³ãã³ã¢ããããã³ïŒâã
ã¹ãâαââãªãã·ã«ãããã¹ããšãŒãã«ãã
é µçŽ åå¿ã«ãŠçæãéé¢ãããNMNã®å®éã«æ
çšã§ãããããã«é µçŽ åå¿ç³»ã«ããããããŒãŒæŽ»
æ§æž¬å®ãADPããããŒãŒåºè³ªçšãªã³ååç©ãã
ã³ãã³ã¢ãããïŒâãã¹ãâαââãªãã·ã«ã
ããã¹ããšãŒãã®ããããäžæåã®å®éã«å©çšã
ãããšãå¯èœã§ããã
ãã詊æäžã«éé¢ããŠããATPã®å®éãããã
ãŒãŒãADPããã³ãããŒãŒåºè³ªçšãªã³ååç©ã«
ããé µçŽ åå¿ã«ãŠADPããªã³é žåãããŠçæã
éé¢ãããATPã®å®éãNMNã®å®éãNMNã
ããã¹ããªã©ãŒãŒããã³ãã³ã¢ããããã³ïŒâã
ã¹ãâαââãªãã·ã«ãããã¹ããšãŒãã«ãã
é µçŽ åå¿ã«ãŠçæãéé¢ãããNMNã®å®éã«æ
çšã§ãããããã«é µçŽ åå¿ç³»ã«ããããããŒãŒæŽ»
æ§æž¬å®ãADPããããŒãŒåºè³ªçšãªã³ååç©ãã
ã³ãã³ã¢ãããïŒâãã¹ãâαââãªãã·ã«ã
ããã¹ããšãŒãã®ããããäžæåã®å®éã«å©çšã
ãããšãå¯èœã§ããã
ãããã®å®éã«ãããŠãATPãŸãã¯NMNã®
ïŒã¢ã«æ¯ã«å¯ŸããŠæ¥µããŠé«ã¢ã«æ¯ã®çæç©è³ªã圢
æãããããããATPãŸãã¯NMNã®é«æåºŠæž¬
宿³ãšããŠæçšã§ããã
ïŒã¢ã«æ¯ã«å¯ŸããŠæ¥µããŠé«ã¢ã«æ¯ã®çæç©è³ªã圢
æãããããããATPãŸãã¯NMNã®é«æåºŠæž¬
宿³ãšããŠæçšã§ããã
次ã«å®æœäŸãæããŠæ¬çºæãå
·äœçã«èª¬æãã
ããããã«ããæ¬çºæãéå®ãããã®ã§ã¯ãªãã
ããããã«ããæ¬çºæãéå®ãããã®ã§ã¯ãªãã
宿œäŸ ïŒ
ïŒè©Šè¬ïŒ
(1) 50ïœïŒ TrisâHClç·©è¡æ¶²PH7.5
(2) ïŒïœïŒ MgCl2
(3) ïŒïœïŒ NMN
(4) NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒïŒã€
ãŒã¹ãç±æ¥ïŒ18UïŒmlïŒ (1) 100ïœïŒ ãªã³é žç·©è¡æ¶²PH8.0 (2) ã¢ã«ã³ãŒã«ããããã²ããŒãŒïŒã€ãŒã¹ãç±
æ¥ïŒ240UïŒmlïŒ (3) 0.01ïŒ NTB (4) ãžã¢ãã©ãŒãŒïŒããã«ã¹ã¡ã¬ããªãŠã ç±
æ¥ïŒ240UïŒmlïŒ (5) 0.8M ãšã¿ããŒã« (6) 0.1ïŒ ããªãã³ïŒžâ100 ïŒæäœïŒ 詊è¬0.1mlã«0.5ã2.5ÎŒMã®ATP10ÎŒãæ·»å
ãã37âã§15åéåå¿ãè¡ãªããåå¿åŸ37âã«äº
åå æž©ããåŸã詊è¬ã0.1mlæ·»å ããæ£ç¢ºã«10
åéåå¿ãè¡ãªãã2.8mlã®0.1NHClãæ·»å ãã
åå¿ã忢ããã®ã¡ã550nïœã«ãŠæ¯è²å®éãã
ããïŒçµæïŒç¬¬ïŒå³ã®éãã§ãã€ãŠãATPã®æ¿åºŠ
ãš550nïœã«ãããåžå 床ã®éã«ããããŠè¯å¥œãª
çŽç·çé¢ä¿ãåŸãããã
ãŒã¹ãç±æ¥ïŒ18UïŒmlïŒ (1) 100ïœïŒ ãªã³é žç·©è¡æ¶²PH8.0 (2) ã¢ã«ã³ãŒã«ããããã²ããŒãŒïŒã€ãŒã¹ãç±
æ¥ïŒ240UïŒmlïŒ (3) 0.01ïŒ NTB (4) ãžã¢ãã©ãŒãŒïŒããã«ã¹ã¡ã¬ããªãŠã ç±
æ¥ïŒ240UïŒmlïŒ (5) 0.8M ãšã¿ããŒã« (6) 0.1ïŒ ããªãã³ïŒžâ100 ïŒæäœïŒ 詊è¬0.1mlã«0.5ã2.5ÎŒMã®ATP10ÎŒãæ·»å
ãã37âã§15åéåå¿ãè¡ãªããåå¿åŸ37âã«äº
åå æž©ããåŸã詊è¬ã0.1mlæ·»å ããæ£ç¢ºã«10
åéåå¿ãè¡ãªãã2.8mlã®0.1NHClãæ·»å ãã
åå¿ã忢ããã®ã¡ã550nïœã«ãŠæ¯è²å®éãã
ããïŒçµæïŒç¬¬ïŒå³ã®éãã§ãã€ãŠãATPã®æ¿åºŠ
ãš550nïœã«ãããåžå 床ã®éã«ããããŠè¯å¥œãª
çŽç·çé¢ä¿ãåŸãããã
宿œäŸ ïŒ
宿œäŸïŒã§çšãã詊è¬ã®ãã¡ãã¢ã«ã³ãŒã«ãã
ããã²ããŒãŒããã³0.8Mãšã¿ããŒã«ã®ä»£ããã«ã
ã°ãªã»ããŒã«âïŒâãã¹ããšãŒãããããã²ããŒ
ãŒïŒãŠãµã®çèç±æ¥ãEC.1.1.1.8ïŒ300UïŒmlãã
ã³10ïœïŒã°ãªã»ããŒã«â3â²âãã¹ããšãŒããçšã
ãŠã以äžå®æœäŸïŒãšåæ§ã®åå¿ãè¡ãªã€ããçµæ
ã¯ç¬¬ïŒå³ã®éãã§ãã€ãŠã宿œäŸïŒãšåæ§ã«ã
ATPãš550nïœã«ãããåžå 床ã®éã«ããããŠè¯
奜ãªçŽç·é¢ä¿ãåŸãããã
ããã²ããŒãŒããã³0.8Mãšã¿ããŒã«ã®ä»£ããã«ã
ã°ãªã»ããŒã«âïŒâãã¹ããšãŒãããããã²ããŒ
ãŒïŒãŠãµã®çèç±æ¥ãEC.1.1.1.8ïŒ300UïŒmlãã
ã³10ïœïŒã°ãªã»ããŒã«â3â²âãã¹ããšãŒããçšã
ãŠã以äžå®æœäŸïŒãšåæ§ã®åå¿ãè¡ãªã€ããçµæ
ã¯ç¬¬ïŒå³ã®éãã§ãã€ãŠã宿œäŸïŒãšåæ§ã«ã
ATPãš550nïœã«ãããåžå 床ã®éã«ããããŠè¯
奜ãªçŽç·é¢ä¿ãåŸãããã
宿œäŸ ïŒ
宿œäŸïŒã«ããã詊è¬ã®ãã¡ãã¢ã«ã³ãŒã«ã
ãããã²ããŒãŒããã³0.8Mãšã¿ããŒã«ãã°ãªã»
ããŒã«ããããã²ããŒãŒïŒããã«ã¹ã»ã¡ã¬ããªãŠ
ã ç±æ¥ãEC.1.1.1.6ïŒ260UïŒmlããã³1Mã°ãªã»
ããŒã«ãçšããŠã以äžå®æœäŸïŒãšåæ§ã®åå¿ãè¡
ãªã€ããçµæã¯ç¬¬ïŒå³ã«ç€ºãããéãã§ããã
ãããã²ããŒãŒããã³0.8Mãšã¿ããŒã«ãã°ãªã»
ããŒã«ããããã²ããŒãŒïŒããã«ã¹ã»ã¡ã¬ããªãŠ
ã ç±æ¥ãEC.1.1.1.6ïŒ260UïŒmlããã³1Mã°ãªã»
ããŒã«ãçšããŠã以äžå®æœäŸïŒãšåæ§ã®åå¿ãè¡
ãªã€ããçµæã¯ç¬¬ïŒå³ã«ç€ºãããéãã§ããã
宿œäŸ ïŒ
ïŒè©Šè¬ïŒ
(1) 50ïœïŒ TrisâHClç·©è¡æ¶²PH7.5
(2) ïŒïœïŒ MgCl2
(3) ïŒïœïŒ ATP
(4) NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒïŒã€
ãŒã¹ãç±æ¥ïŒ18UïŒmlïŒ (1) 100ïœïŒ ãªã³é žç·©è¡æ¶² PH8.0 (2) ã¢ã«ã³ãŒã«ããããã²ããŒãŒïŒã€ãŒã¹ãç±
æ¥ïŒ240UïŒmlïŒ (3) 0.01ïŒ NTB (4) ãžã¢ãã©ãŒãŒïŒããã«ã¹ã¡ã¬ããªãŠã ç±
æ¥ïŒ240UïŒmlïŒ (5) 0.8M ãšã¿ããŒã« (6) 0.1ïŒ ããªãã³ïŒžâ100 ïŒæäœïŒ 詊è¬0.1mlã«0.5ã2.5ÎŒMã®NMN10ÎŒãæ·»å
ãã37âã§15åéåå¿ãè¡ãªããåå¿åŸ37âã«äº
åå æž©ããåŸã詊è¬ã0.1mlæ·»å ããæ£ç¢ºã«10
åéåå¿ãè¡ãªãã2.8mlã®0.1NHClãæ·»å ãã
åå¿ã忢ããã®ã¡ã550nïœã«ãŠæ¯è²å®éããã
ãŒã¹ãç±æ¥ïŒ18UïŒmlïŒ (1) 100ïœïŒ ãªã³é žç·©è¡æ¶² PH8.0 (2) ã¢ã«ã³ãŒã«ããããã²ããŒãŒïŒã€ãŒã¹ãç±
æ¥ïŒ240UïŒmlïŒ (3) 0.01ïŒ NTB (4) ãžã¢ãã©ãŒãŒïŒããã«ã¹ã¡ã¬ããªãŠã ç±
æ¥ïŒ240UïŒmlïŒ (5) 0.8M ãšã¿ããŒã« (6) 0.1ïŒ ããªãã³ïŒžâ100 ïŒæäœïŒ 詊è¬0.1mlã«0.5ã2.5ÎŒMã®NMN10ÎŒãæ·»å
ãã37âã§15åéåå¿ãè¡ãªããåå¿åŸ37âã«äº
åå æž©ããåŸã詊è¬ã0.1mlæ·»å ããæ£ç¢ºã«10
åéåå¿ãè¡ãªãã2.8mlã®0.1NHClãæ·»å ãã
åå¿ã忢ããã®ã¡ã550nïœã«ãŠæ¯è²å®éããã
ïŒçµæïŒ
第ïŒå³ã«ç€ºãããéãã§ãNMNã®æ¿åºŠãš550n
ïœã«ãããåžå 床ã®éã«ããããŠè¯å¥œãªçŽç·çé¢
ä¿ãåŸãããã
ïœã«ãããåžå 床ã®éã«ããããŠè¯å¥œãªçŽç·çé¢
ä¿ãåŸãããã
宿œäŸ ïŒ
ïŒåå¿æ¶²ïŒ
(1) 50ïœïŒ TrisâHClç·©è¡æ¶²PH7.5
(2) 20ïœïŒ MgCl2
(3) ã¢ã»ããŒããããŒãŒïŒå€§è
žèç±æ¥ã8UïŒmlïŒ
(4) 10ïœïŒ MgCl2
(5) ïŒïœïŒ NMN
(6) NMNã¢ããã«ãã©ã¹ã¹ããšã©ãŒãŒïŒ18UïŒ
mlïŒ ïŒæäœïŒ åå¿æ¶²0.2mlãå°è©Šéšç®¡ã«ãšããåæ¿åºŠã®ã¢
ã»ãã«ãã¹ããšãŒã溶液10ÎŒãæ·»å ãã37âã§
20ååå¿ãè¡ãªã€ãã®ã¡ã宿œäŸïŒã«ç€ºãã詊è¬
ã0.2mlå ãã10åéåå¿ããã®ã¡ã1.8mlã®
0.1NHCl溶液ãå ããŠåå¿ã忢ããã®ã¡550n
ïœã§æ¯è²å®éããçµæã第ïŒå³ã«ç€ºãåŠãè¯å¥œãª
çŽç·ãåŸãããã
mlïŒ ïŒæäœïŒ åå¿æ¶²0.2mlãå°è©Šéšç®¡ã«ãšããåæ¿åºŠã®ã¢
ã»ãã«ãã¹ããšãŒã溶液10ÎŒãæ·»å ãã37âã§
20ååå¿ãè¡ãªã€ãã®ã¡ã宿œäŸïŒã«ç€ºãã詊è¬
ã0.2mlå ãã10åéåå¿ããã®ã¡ã1.8mlã®
0.1NHCl溶液ãå ããŠåå¿ã忢ããã®ã¡550n
ïœã§æ¯è²å®éããçµæã第ïŒå³ã«ç€ºãåŠãè¯å¥œãª
çŽç·ãåŸãããã
第ïŒå³ã¯æç€ºåå¿ç³»ã«ã¢ã«ã³ãŒã«ããããã²ã
ãŒãŒããšã¿ããŒã«ããžã¢ãã©ãŒãŒãããã©ãŸãªãŠ
ã å¡©ãçšããŠãªãATPã®æ€éæ²ç·ã瀺ãã第ïŒ
å³ã¯æç€ºåå¿ã«ã°ãªã»ããŒã«âïŒâãã¹ããšãŒã
ããããã²ããŒãŒãã°ãªã»ããŒã«â3â²âãã¹ããš
ãŒãããžã¢ãã©ãŒãŒãããã©ãŸãªãŠã å¡©ãçšããŠ
ãªãATPã®æ€éæ²ç·ã瀺ãã第ïŒå³ã¯æç€ºåå¿
ç³»ã«ã°ãªã»ããŒã«ããããã²ããŒãŒãã°ãªã»ããŒ
ã«ããžã¢ãã©ãŒãŒãããã©ãŸãªãŠã å¡©ãçšããŠãª
ãATPã®æ€éæ²ç·ã瀺ãã第ïŒå³ã¯æç€ºåå¿ç³»
ã«ã¢ã«ã³ãŒã«ããããã²ããŒãŒããšã¿ããŒã«ããž
ã¢ãã©ãŒãŒãããã©ãŸãªãŠã å¡©ãçšããŠãªã
NMNã®æ€éæ²ç·ã瀺ãã第ïŒå³ã¯æ¬çºæã«ãã
ã¢ã»ãã«ãã¹ããšãŒãã®æ€éæ²ç·ã瀺ãã
ãŒãŒããšã¿ããŒã«ããžã¢ãã©ãŒãŒãããã©ãŸãªãŠ
ã å¡©ãçšããŠãªãATPã®æ€éæ²ç·ã瀺ãã第ïŒ
å³ã¯æç€ºåå¿ã«ã°ãªã»ããŒã«âïŒâãã¹ããšãŒã
ããããã²ããŒãŒãã°ãªã»ããŒã«â3â²âãã¹ããš
ãŒãããžã¢ãã©ãŒãŒãããã©ãŸãªãŠã å¡©ãçšããŠ
ãªãATPã®æ€éæ²ç·ã瀺ãã第ïŒå³ã¯æç€ºåå¿
ç³»ã«ã°ãªã»ããŒã«ããããã²ããŒãŒãã°ãªã»ããŒ
ã«ããžã¢ãã©ãŒãŒãããã©ãŸãªãŠã å¡©ãçšããŠãª
ãATPã®æ€éæ²ç·ã瀺ãã第ïŒå³ã¯æç€ºåå¿ç³»
ã«ã¢ã«ã³ãŒã«ããããã²ããŒãŒããšã¿ããŒã«ããž
ã¢ãã©ãŒãŒãããã©ãŸãªãŠã å¡©ãçšããŠãªã
NMNã®æ€éæ²ç·ã瀺ãã第ïŒå³ã¯æ¬çºæã«ãã
ã¢ã»ãã«ãã¹ããšãŒãã®æ€éæ²ç·ã瀺ãã
Claims (1)
- ãç¹èš±è«æ±ã®ç¯å²ã ïŒ ATPïŒã¢ããã·ã³ããªãã¹ããšãŒãïŒããã³
NMNïŒãã³ãã³é žã¢ããã¢ããã¯ã¬ãªããïŒã®
ããããäžæåã嫿ããè¢«æ€æ¶²ã«ãäž»åå¿ç³»ã«
ãããŠNMNãŸãã¯ATPããã°ãã·ãŠã ã€ãªã³
ã®ååšäžã«NMNã¢ãããªã«ãã©ã³ã¹ããšã©ãŒãŒ
ãäœçšãããŠNADãšãããªã³é žã«å€æãããäž»
åå¿ã®åŸãçããNADããNADïŒïŒ°ïŒãè£é µçŽ
ãšããé žåéå åå¿ç³»ãšãéå åNADïŒïŒ°ïŒãè£
é µçŽ ãšããåå¿ç³»ãšã®çµåãã«ããè£é µçŽ ãµã€ã¯
ãªã³ã°åå¿ããããªããæ¬¡ãã§ãã®ãµã€ã¯ãªã³ã°
åå¿ã«ãããŠçæãŸãã¯æ¶è²»ãããæåãå®éã
ãããšãç¹åŸŽãšããATPããã³NMNã®ããã
ãäžæåã嫿ããè¢«æ€æ¶²äžã®æåã®é«æåºŠæž¬å®
æ³ã ïŒ ATPãããããŒãŒãADPïŒã¢ããã·ã³ãžã
ã¯ã¬ãªããïŒããã³ãããŒãŒåºè³ªçšãªã³ååç©ã®
åå¿ç³»ã«ãã€ãŠçæãããATPã§ããç¹èš±è«æ±
ã®ç¯å²ç¬¬ïŒé èšèŒã®é«æåºŠæž¬å®æ³ã ïŒ ãããŒãŒãADPããã³ãããŒãŒåºè³ªçšãªã³
ååç©ã®åå¿ç³»ããã¯ã¬ã¢ãã³ãããŒãŒãADP
ããã³ã¯ã¬ã¢ãã³ãã¹ããšãŒãã®åå¿ç³»ããã«ã
ãŒããããŒãŒãADPããã³ãã¹ããšããŒã«ãã«
ãã³é žã®åå¿ç³»ãã¢ã»ããŒããããŒãŒãADPã
ã¢ã»ãã«ãã¹ããšãŒãã®åå¿ç³»ãã«ã«ãã¡ãŒãã
ããŒãŒãADPããã³ã«ã«ãã¢ã€ã«ãã¹ããšãŒã
ã®åå¿ç³»ãã¢ã¹ãã«ãã€ããããŒãŒãADPãïŒ
âãã¹ãââã¢ã¹ãã«ãã€ãã®åå¿ç³»ããã¹ã
ã°ãªã»ã¬ã€ããããŒãŒãADPã1.3âãžãã¹ãâ
âã°ãªã»ã¬ã€ãã®åå¿ç³»ãã¢ã«ã®ãã³ãããŒ
ãŒãADPãã¢ã«ã®ãã³ãã¹ããšãŒãã®åå¿ç³»ã
ããªãããŒãŒãADPãADPã®åå¿ç³»ããŸãã¯ã
ã¯ã¬ãªãµã€ãã¢ããã¹ããšãŒããããŒãŒãADPã
ãã¯ã¬ãªãµã€ããžãã¹ããšãŒãã®åå¿ç³»ã§ããç¹
èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®é«æåºŠæž¬å®æ³ã ïŒ NMNãäžèšã«ç€ºãããé µçŽ åå¿ã«ãã€ãŠç
æãããNMNã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒ
ã®é«æåºŠæž¬å®æ³ã ãã³ãã³ã¢ããïŒïŒâãã¹ãâαââãªãã· ã«ãããã¹ããšãŒã âââââââââââââââ NMNãããã¹ããªãªã©ãŒãŒNMNïŒãããªã³é ž ïŒ NADïŒïŒ°ïŒãè£é µçŽ ãšããé žåéå åå¿ç³»
ããå°ãªããšãNADãæ¶è²»ããŠéå åNADãç
æããåå¿ã圢æããããããã²ããŒãŒããã³ã
ã®åºè³ªã«ããåå¿ç³»ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé
èšèŒã®é«æåºŠæž¬å®æ³ã ïŒ ããããã²ããŒãŒããã³ãã®åºè³ªããã©ã¯ã
ãŒãããããã²ããŒãŒããã³ïŒ¬âã©ã¯ããŒããã¢
ã«ã³ãŒã«ããããã²ããŒãŒããã³ãšã¿ããŒã«ãã°
ãªã»ããŒã«ããããã²ããŒãŒããã³ã°ãªã»ããŒ
ã«ãã°ãªã»ããŒã«âïŒâãã¹ããšãŒãããããã²
ããŒãŒããã³ã°ãªã»ããŒã«âïŒâãã¹ããšãŒãã
ã°ã«ã³ãŒã¹ããããã²ããŒãŒããã³ã°ã«ã³ãŒã¹ã
ãã¬ãŒãããããã²ããŒãŒããã³ïŒ¬âãã¬ãŒãã
ã°ã«ã¿ã¡ã€ãããããã²ããŒãŒããã³ïŒ¬âã°ã«ã¿
ã¡ã€ããïŒâαâããããã·ã¹ããã€ããããã
ã²ããŒãŒããã³ïŒâαâããããã·ã¹ããã€ãã®
ããããã®åå¿ç³»ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èš
èŒã®é«æåºŠæž¬å®æ³ã ïŒ éå åNADïŒïŒ°ïŒãè£é µçŽ ãšããåå¿ç³»ãã
å°ãªããšãéå åNADãæ¶è²»ããŠNADãçæã
ãåå¿ã圢æããé žåéå é µçŽ ããã³ãã®åºè³ªã
ããªãåå¿ç³»ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®
é«æåºŠæž¬å®æ³ã ïŒ å°ãªããšãéå åNADãæ¶è²»ããŠNADãç
æããé žåéå é µçŽ ããã³ãã®åºè³ªããã©ã¯ããŒ
ãããããã²ããŒãŒããã³ãã«ãã³é žãã¢ã«ã³ãŒ
ã«ããããã²ããŒãŒããã³ã¢ã»ãã¢ã«ããããã°
ãªã»ããŒã«ããããã²ããŒãŒããã³ãžããããã·
ã¢ã»ãã³ãã°ãªã»ããŒã«âïŒâãã¹ããšãŒããã
ããã²ããŒãŒããã³ã·ããããã·ã¢ã»ãã³ãã¹ã
ãšãŒãããã¬ãŒãããããã²ããŒãŒããã³ãªã®ã¶
ãé ¢é žãïŒâαâããããã·ã¹ããã€ããããã
ã²ããŒãŒããã³ïŒâã±ãã¹ããã€ãã®ããããã®
åå¿ç³»ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®é«æåºŠ
æž¬å®æ³ã ïŒ å°ãªããšãéå åNADãæ¶è²»ããŠNADãç
æããåå¿ã圢æããé žåéå é µçŽ ããã³ãã®åº
質ããéå åNADïŒïŒ°ïŒïŒå容äœé žåéå é µçŽ ã
ãã³ãã®å容äœã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒ
ã®é«æåºŠæž¬å®æ³ã ïŒïŒ éå åNADïŒïŒ°ïŒïŒå容äœé žåéå é µçŽ ã
ãã³ãã®å容äœããéå åNADïŒïŒ°ïŒããããã²
ããŒãŒããã³ãã©ãã³é¡ãããã³é¡ãïŒïŒïŒâãž
ã¯ããããšããŒã«ã€ã³ãããšããŒã«ãããšãªã·ã¢
ã³åå¡©ãããã©ãŸãªãŠã å¡©ãããã¯ããŒã ãŸã
ã¯é žçŽ ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®é«æåºŠ
æž¬å®æ³ã ïŒïŒ éå åNADïŒïŒ°ïŒïŒå容äœé žåéå åé µçŽ
ããã³ãã®åºè³ªãããžã¢ãã©ãŒãŒããã³ããã©ãŸ
ãªãŠã å¡©ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®é«æ
åºŠæž¬å®æ³ã ïŒïŒ éå åNADïŒïŒ°ïŒãè£é µçŽ ãšããåå¿ç³»
ããé»åäŒéç©è³ªããã³ããã©ãŸãªãŠã å¡©ã§ãã
ç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒã®é«æåºŠæž¬å®æ³ã ïŒïŒ é»åäŒéç©è³ªããããšããžã³âã¡ã¿ãµã«ã
ãšãŒããã¡ã«ããŒã©ã»ãã«ãŒãŸãã¯ããã·ã¢ãã³
ã§ããç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒïŒé èšèŒã®é«æåºŠæž¬å®
æ³ã ïŒïŒ ãµã€ã¯ãªã³ã°åå¿ã«ãããŠãç颿޻æ§å€ã
æ·»å ããããšãããªãç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒé èšèŒ
ã®é«æåºŠæž¬å®æ³ã ïŒïŒ ç颿޻æ§å€ãéã€ãªã³ç³»ç颿޻æ§å€ã§ãã
ç¹èš±è«æ±ã®ç¯å²ç¬¬ïŒïŒé èšèŒã®é«æåºŠæž¬å®æ³ã
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4120283A JPS59166099A (ja) | 1983-03-11 | 1983-03-11 | ïœïœãŸãã¯ïœïœïœã®é«æåºŠæž¬å®æ³ |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP4120283A JPS59166099A (ja) | 1983-03-11 | 1983-03-11 | ïœïœãŸãã¯ïœïœïœã®é«æåºŠæž¬å®æ³ |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS59166099A JPS59166099A (ja) | 1984-09-19 |
| JPH047200B2 true JPH047200B2 (ja) | 1992-02-10 |
Family
ID=12601828
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP4120283A Granted JPS59166099A (ja) | 1983-03-11 | 1983-03-11 | ïœïœãŸãã¯ïœïœïœã®é«æåºŠæž¬å®æ³ |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPS59166099A (ja) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2003044222A1 (fr) | 2001-11-21 | 2003-05-30 | Unitika Ltd. | Procede de mesure atp permettant une evaluation visuelle et reactif utilise a cet effet |
| JP5570731B2 (ja) | 2008-02-28 | 2014-08-13 | æåæãã¡ãŒãæ ªåŒäŒç€Ÿ | ãããªã³é žã®æž¬å®æ¹æ³ |
| JP6917239B2 (ja) * | 2017-08-03 | 2021-08-11 | æåæãã¡ãŒãæ ªåŒäŒç€Ÿ | ã¿ã³ãã¯è³ªããã³ãã®è£œé æ¹æ³ãã¿ã³ãã¯è³ªçµæç©ãªãã³ã«ãããçšããïœïœïœã®æž¬å®æ¹æ³ããã³æž¬å®çµæç© |
| WO2021241446A1 (ja) | 2020-05-25 | 2021-12-02 | æšªæ²³é»æ©æ ªåŒäŒç€Ÿ | æ€äœäžã®æšçååã®æ€åºæ¹æ³ãåã³æšçå忀åºããã |
-
1983
- 1983-03-11 JP JP4120283A patent/JPS59166099A/ja active Granted
Also Published As
| Publication number | Publication date |
|---|---|
| JPS59166099A (ja) | 1984-09-19 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP0034213B1 (en) | A stabilized aqueous coenzyme solution for use in a clinical assay, a method of stabilizing a labile coenzyme in an aqueous clinical assay solution and a kit for use in said clinical assay | |
| US4242446A (en) | Method for determining a substance in a biological fluid and reagent combination for use in the method | |
| US4241179A (en) | Method for determining a transaminase in a biological fluid and reagent combination for use in the method | |
| Kirstein et al. | Highly sensitive enzyme thermistor determination of ADP and ATP by multiple recycling enzyme systems | |
| JPH047200B2 (ja) | ||
| US7122333B2 (en) | Method and reagent for visually measuring ATP | |
| JPH0220239B2 (ja) | ||
| US3985621A (en) | Stopping agents for enzyme reactions of dehydrogenase systems | |
| EP0207493B1 (en) | Method of terminating isocitrate dehydrogenase reaction | |
| US4956276A (en) | Analytical method of determining a reduced co-enzyme | |
| US10472665B2 (en) | Measuring method and composition using kinase | |
| US5081014A (en) | Method of measuring a co-enzyme | |
| JP2818696B2 (ja) | ïœïœïœãããŒãŒãçšããé«æåºŠå®éæ³ | |
| WO2012050536A1 (en) | Method of the adenosine diphosphate quantitative determination | |
| JPH0675515B2 (ja) | ã¢ã³ã¢ãã¢ãåå¿çæç©ãšããçäœç©è³ªã®å®éæ¹æ³ | |
| US3838010A (en) | Photometric method for determining lactic acid dehydrogenase or glucose-6-phosphate dehydrogenase | |
| JPH0412120B2 (ja) | ||
| JP3586737B2 (ja) | çäœç©è³ªã®æž¬å®æ¹æ³ | |
| JP3566976B2 (ja) | çäœæåäžã®æž¬å®å¯Ÿè±¡ã®æž¬å®æ¹æ³ | |
| JP2001252095A (ja) | 埮éæåã®æž¬å®æ³åã³ããã«çšããçµæç© | |
| JP5633669B2 (ja) | ïœïœã®æž¬å®æ¹æ³ããã³ïœïœïœæž¬å®çšããã | |
| JPH0560920B2 (ja) | ||
| Grassl | Guanosine-5â²-triphosphate and inosine-5â²-triphosphate | |
| JP3034979B2 (ja) | ã°ãªã»ããŒã«ããžããããã·ã¢ã»ãã³ãŸãã¯ïœâã°ãªã»ãã¢ã«ãããã®é«æåºŠå®éæ³ããã³é«æåºŠå®éçšçµæç© | |
| JPH07108239B2 (ja) | ãã«ãã³é žã®å®éæ¹æ³åã³è©²æ¹æ³ãå©çšããçäœæåã®å®éæ¹æ³ |