WO2001007638A3 - Homogeneous fluorescence method for assaying structural modifications of biomolecules - Google Patents

Homogeneous fluorescence method for assaying structural modifications of biomolecules Download PDF

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Publication number
WO2001007638A3
WO2001007638A3 PCT/US2000/040495 US0040495W WO0107638A3 WO 2001007638 A3 WO2001007638 A3 WO 2001007638A3 US 0040495 W US0040495 W US 0040495W WO 0107638 A3 WO0107638 A3 WO 0107638A3
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WO
WIPO (PCT)
Prior art keywords
substrates
substrate
dyes
labeled
biomolecules
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.)
Ceased
Application number
PCT/US2000/040495
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French (fr)
Other versions
WO2001007638A2 (en
Inventor
Donald K Ii Blumenthal
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
University of Utah Research Foundation Inc
Original Assignee
University of Utah Research Foundation Inc
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by University of Utah Research Foundation Inc filed Critical University of Utah Research Foundation Inc
Priority to AU76271/00A priority Critical patent/AU7627100A/en
Priority to CA002380238A priority patent/CA2380238A1/en
Priority to EP00965572A priority patent/EP1206699A4/en
Publication of WO2001007638A2 publication Critical patent/WO2001007638A2/en
Publication of WO2001007638A3 publication Critical patent/WO2001007638A3/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/536—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase
    • G01N33/542—Immunoassay; Biospecific binding assay; Materials therefor with immune complex formed in liquid phase with steric inhibition or signal modification, e.g. fluorescent quenching
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6813—Hybridisation assays
    • C12Q1/6816—Hybridisation assays characterised by the detection means
    • C12Q1/6818—Hybridisation assays characterised by the detection means involving interaction of two or more labels, e.g. resonant energy transfer

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Zoology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Molecular Biology (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Analytical Chemistry (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Urology & Nephrology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Hematology (AREA)
  • Cell Biology (AREA)
  • Food Science & Technology (AREA)
  • Medicinal Chemistry (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

Double-labeled protein biomolecular substrates (70) and methods for the homogenous assay of processes which include covalent modification of the substrates (70) to form a detectable species are described. The biomolecular substrates (70) of the instant invention are labeled at two positions (80, 90) with two fluorescent dyes or with a fluorescent dye and a nonfluorescent dye. The two labeling dyes of the unmodified substrate (70) stack (95), thereby quenching the substrate's fluorescence. Upon covalent modification of the double-labeled substrate (70), however, the intramolecularly stacked dyes (95) dissociate and the fluorescence changes markedly. Examples are described for the preparation and use of substrates (70) for phosphorylation assays. Methods of invention do not require separation of the modified and unmodified substrates (70), nor do they require other special reagents or radioactive materials. Therefore the substrates can be used for monitoring intracellular processes of living cells.
PCT/US2000/040495 1999-07-27 2000-07-27 Homogeneous fluorescence method for assaying structural modifications of biomolecules Ceased WO2001007638A2 (en)

Priority Applications (3)

Application Number Priority Date Filing Date Title
AU76271/00A AU7627100A (en) 1999-07-27 2000-07-27 Homogeneous fluorescence method for assaying structural modifications of biomolecules
CA002380238A CA2380238A1 (en) 1999-07-27 2000-07-27 Homogeneous fluorescence method for assaying structural modifications of biomolecules
EP00965572A EP1206699A4 (en) 1999-07-27 2000-07-27 HOMOGENEOUS FLUORESCENCE METHOD FOR DETERMINING STRUCTURAL MODIFICATIONS OF BIOMOLECULES

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US14575599P 1999-07-27 1999-07-27
US60/145,755 1999-07-27

Publications (2)

Publication Number Publication Date
WO2001007638A2 WO2001007638A2 (en) 2001-02-01
WO2001007638A3 true WO2001007638A3 (en) 2001-08-16

Family

ID=22514394

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2000/040495 Ceased WO2001007638A2 (en) 1999-07-27 2000-07-27 Homogeneous fluorescence method for assaying structural modifications of biomolecules

Country Status (4)

Country Link
EP (1) EP1206699A4 (en)
AU (1) AU7627100A (en)
CA (1) CA2380238A1 (en)
WO (1) WO2001007638A2 (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003106703A2 (en) * 2002-06-14 2003-12-24 Pierce Biotechnology, Inc. Homogenous assay for enzymatic activity
WO2005071095A2 (en) * 2004-01-16 2005-08-04 Applera Corporation Fluorogenic kinase assays and substrates for kinases and phosphatases
EP1759015A2 (en) * 2004-06-21 2007-03-07 Applera Corporation, Applied Biosystems Group Fluorogenic enzyme assays and substrates for kinases and phosphatases
EP1634603A1 (en) * 2004-08-26 2006-03-15 Eberhard-Karls-Universität Tübingen Universitätsklinikum Treatment of transformed or infected biologic Cells
WO2010060972A1 (en) * 2008-11-26 2010-06-03 Julius-Maximilians-Universitaet Wuerzburg Erk1/2 phosphorylation site specific antibody

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4777128A (en) * 1986-05-27 1988-10-11 Ethigen Corporation Fluorescence immunoassay involving energy transfer between two fluorophores
US5654419A (en) * 1994-02-01 1997-08-05 The Regents Of The University Of California Fluorescent labels and their use in separations
US5723340A (en) * 1993-09-30 1998-03-03 Avl Medical Instruments Ag Optical indicator for determining the activity of an ion in a sample
US5869255A (en) * 1994-02-01 1999-02-09 The Regents Of The University Of California Probes labeled with energy transfer couples dyes exemplified with DNA fragment analysis
FR2768817A1 (en) * 1997-09-19 1999-03-26 Cis Bio Int HOMOGENEOUS METHOD FOR THE DETECTION AND/OR DETERMINATION OF THE PHOSPHORYLATING ACTIVITY OF A BIOLOGICAL MATERIAL

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DK0537270T3 (en) * 1990-07-02 1999-06-07 Univ California Detection of analytes by fluorescence energy transfer
ATE428801T1 (en) * 1996-06-04 2009-05-15 Univ Utah Res Found MONITORING HYBRIDIZATION DURING PCR

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4777128A (en) * 1986-05-27 1988-10-11 Ethigen Corporation Fluorescence immunoassay involving energy transfer between two fluorophores
US5723340A (en) * 1993-09-30 1998-03-03 Avl Medical Instruments Ag Optical indicator for determining the activity of an ion in a sample
US5654419A (en) * 1994-02-01 1997-08-05 The Regents Of The University Of California Fluorescent labels and their use in separations
US5707804A (en) * 1994-02-01 1998-01-13 The Regents Of The University Of California Primers labeled with energy transfer coupled dyes for DNA sequencing
US5869255A (en) * 1994-02-01 1999-02-09 The Regents Of The University Of California Probes labeled with energy transfer couples dyes exemplified with DNA fragment analysis
FR2768817A1 (en) * 1997-09-19 1999-03-26 Cis Bio Int HOMOGENEOUS METHOD FOR THE DETECTION AND/OR DETERMINATION OF THE PHOSPHORYLATING ACTIVITY OF A BIOLOGICAL MATERIAL

Non-Patent Citations (13)

* Cited by examiner, † Cited by third party
Title
A.-P. WEI ET AL.: "Antibody-mediated fluorescence enhancement based on shifting the intramolecular dimer/monomer equilibrium of fluorescent dyes", ANALYTICAL CHEMISTRY, vol. 66, no. 9, 1 May 1994 (1994-05-01), pages 1500 - 1506, XP002937508 *
B. BISPISA ET AL.: "Intramolecular electronic energy transfer between naphthalene and protoporphyrin chromophores bound to sequential peptides", MACROMOLECULES, vol. 27, no. 26, 19 December 1994 (1994-12-19), pages 7800 - 7808, XP002937507 *
C.G. DOS REMEDIOS ET AL.: "Fluorescence resonance energy transfer spectroscopy is a reliable 'ruler' for measuring structural changes in proteins", JOURNAL OF STRUCTURAL BIOLOGY, vol. 115, 1995, pages 175 - 185, XP002937501 *
D.A. PEARCE ET AL.: "Peptidyl chemosensors incorporating a FRET mechanism for detection of Ni(II)", BIOORGANIC & MEDICINAL CHEMISTRY LETTERS, vol. 8, no. 15, 4 August 1998 (1998-08-04), pages 1963 - 1968, XP002937503 *
G. HARNISCHFEGER: "Effect of chemical modification of amino groups by fluorescamine on partial reactions of photosynthesis", BIOCHIMICA ET BIOPHYSICA ACTA, vol. 503, no. 3, 7 September 1978 (1978-09-07), pages 473 - 479, XP002937510 *
J. JU ET AL.: "Cassette labeling for facile construction of energy transfer fluorescent primers", NUCLEIC ACIDS RESEARCH, vol. 24, no. 6, 15 March 1996 (1996-03-15), pages 1144 - 1148, XP002937505 *
K.B. LEE ET AL.: "A new approach to assay endo-type carbohydrases: Bifluorescent-labeled substrates for glycoamides and ceramide glycanases", ANALYTICAL BIOCHEMISTRY, vol. 230, no. 1, 1 September 1995 (1995-09-01), pages 31 - 36, XP002937512 *
K.M. PARKHURST ET AL.: "Kinetic studies by fluorescence resonance energy transfer employing a double-labeled oligonucleotide: Hybridization to the oligonucleotide complement and to single-stranded DNA", BIOCHEMISTRY, vol. 34, no. 1, 10 January 1995 (1995-01-10), pages 285 - 292, XP002937506 *
KOJI MUTSUOKA ET AL.: "A bi-fluorescence-labeled substrate for ceramide glycanase based on fluorescence energy transfer", CARBOHYDRATE RESEARCH, vol. 276, no. 1, 16 October 1995 (1995-10-16), pages 31 - 42, XP002937511 *
L.A. MARCAURELLE ET AL.: "Direct incorporation of unprotected ketone groups into peptides during solid-phase synthesis: Application to the one-step modification of peptides with two different biophysical probes for FRET", TETRAHEDRON LETTERS, vol. 39, no. 40, 1 October 1998 (1998-10-01), pages 7279 - 7282, XP002937502 *
M.P. LILLO ET AL.: "Design and characterization of a multisite fluorescence energy-transfer system for protein folding studies: A seady-state and time-resolved study of yeast phosphoglycerate kinase", BIOCHEMISTRY, vol. 36, no. 37, 16 September 1997 (1997-09-16), pages 11261 - 11272, XP002937504 *
See also references of EP1206699A4 *
ZHUJUN ZHANG ET AL.: "Amylase substrate based on fluorescence energy transfer", ANALYTICA CHIMICA ACTA, vol. 236, no. 2, 17 September 1990 (1990-09-17), pages 251 - 256, XP002937509 *

Also Published As

Publication number Publication date
CA2380238A1 (en) 2001-02-01
EP1206699A4 (en) 2005-01-19
AU7627100A (en) 2001-02-13
WO2001007638A2 (en) 2001-02-01
EP1206699A2 (en) 2002-05-22

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