WO2008140060A1 - 活性化プロテアーゼインジケーター - Google Patents

活性化プロテアーゼインジケーター Download PDF

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Publication number
WO2008140060A1
WO2008140060A1 PCT/JP2008/058669 JP2008058669W WO2008140060A1 WO 2008140060 A1 WO2008140060 A1 WO 2008140060A1 JP 2008058669 W JP2008058669 W JP 2008058669W WO 2008140060 A1 WO2008140060 A1 WO 2008140060A1
Authority
WO
WIPO (PCT)
Prior art keywords
protease
luc
luciferase
activated
linked
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/JP2008/058669
Other languages
English (en)
French (fr)
Inventor
Yoshio Umezawa
Takeaki Ozawa
Akira Kanno
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.)
Kagawa University NUC
University of Tokyo NUC
National Institute of Natural Sciences
Original Assignee
Kagawa University NUC
University of Tokyo NUC
National Institute of Natural Sciences
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 Kagawa University NUC, University of Tokyo NUC, National Institute of Natural Sciences filed Critical Kagawa University NUC
Priority to EP08752552A priority Critical patent/EP2154159A4/en
Priority to US12/451,341 priority patent/US8658777B2/en
Priority to JP2008548903A priority patent/JP4491516B2/ja
Publication of WO2008140060A1 publication Critical patent/WO2008140060A1/ja
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

Links

Classifications

    • 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
    • C12Q1/34—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase
    • C12Q1/37—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving peptidase or proteinase
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0069—Oxidoreductases (1.) acting on single donors with incorporation of molecular oxygen, i.e. oxygenases (1.13)
    • 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
    • C12Q1/66—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving luciferase
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/50—Fusion polypeptide containing protease site
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2500/00—Screening for compounds of potential therapeutic value
    • G01N2500/04—Screening involving studying the effect of compounds C directly on molecule A (e.g. C are potential ligands for a receptor A, or potential substrates for an enzyme A)

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Genetics & Genomics (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Physics & Mathematics (AREA)
  • Analytical Chemistry (AREA)
  • Biophysics (AREA)
  • Immunology (AREA)
  • Medicinal Chemistry (AREA)
  • Biomedical Technology (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

本発明は、活性化プロテアーゼの検出、及び生細胞内におけるプロテアーゼ活性化の非侵襲的・高感度・リアルタイム検出を可能とする新しい手段を提供することを課題とする。本発明の活性化プロテアーゼの検出方法においては、ルシフェラーゼのCハーフ断片（Luc-C）とルシフェラーゼのNハーフ断片（Luc-N）とがプロテアーゼ基質ペプチドによって連結された環状のインジケーターをin vitroのアッセイ系または細胞内に導入する。プロテアーゼによる基質ペプチドの切断によって、Luc-NとLuc-Cが活性化ルシフェラーゼを再構成するため、このルシフェラーゼの発光シグナルを測定することによって、活性化プロテアーゼが検出可能となる。
PCT/JP2008/058669 2007-05-09 2008-05-09 活性化プロテアーゼインジケーター Ceased WO2008140060A1 (ja)

Priority Applications (3)

Application Number Priority Date Filing Date Title
EP08752552A EP2154159A4 (en) 2007-05-09 2008-05-09 INDICATOR FOR ACTIVATED PROTEASE
US12/451,341 US8658777B2 (en) 2007-05-09 2008-05-09 Activated protease indicator
JP2008548903A JP4491516B2 (ja) 2007-05-09 2008-05-09 活性化プロテアーゼインジケーター

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2007124924 2007-05-09
JP2007-124924 2007-05-09

Publications (1)

Publication Number Publication Date
WO2008140060A1 true WO2008140060A1 (ja) 2008-11-20

Family

ID=40002254

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2008/058669 Ceased WO2008140060A1 (ja) 2007-05-09 2008-05-09 活性化プロテアーゼインジケーター

Country Status (4)

Country Link
US (1) US8658777B2 (ja)
EP (1) EP2154159A4 (ja)
JP (1) JP4491516B2 (ja)
WO (1) WO2008140060A1 (ja)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2015530114A (ja) * 2012-09-26 2015-10-15 プロメガ コーポレイションＰｒｏｍｅｇａ Ｃｏｒｐｏｒａｔｉｏｎ リアルタイムモニタリング

Families Citing this family (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2005038029A2 (en) 2003-10-10 2005-04-28 Promega Corporation Luciferase biosensor
EP2004813B1 (en) 2006-04-03 2015-06-03 Promega Corporation Permuted and nonpermuted luciferase biosensors binding cyclic nucleotides
EP2281046B1 (en) 2008-05-19 2012-01-25 Promega Corporation LUCIFERASE BIOSENSORS FOR cAMP
US9290794B2 (en) * 2010-05-11 2016-03-22 Promega Corporation Mutant protease biosensors with enhanced detection characteristics
EP2569423B1 (en) 2010-05-11 2015-12-02 Promega Corporation Mutant protease biosensors with enhanced detection characteristics
EP2776573A1 (en) * 2011-11-11 2014-09-17 Promega Corporation Mutant protease biosensors with enhanced detection characteristics
CA2866595A1 (en) * 2012-03-07 2013-09-12 Merz Pharma Gmbh & Co. Kgaa Means and methods for determining neurotoxin activity based on a modified luciferase
EP3196303B1 (en) * 2014-07-31 2020-07-22 Transgenic Inc. Inflammation reporter system
US20180327779A1 (en) * 2015-03-03 2018-11-15 Fondazione Telethon (IT/IT) Multiple vector system and uses thereof
CN110093319A (zh) * 2019-05-21 2019-08-06 中国农业科学院上海兽医研究所(中国动物卫生与流行病学中心上海分中心) 一种实时指示凋亡水平的稳转细胞系及其构建方法和应用
CN112831479B (zh) * 2021-01-05 2022-10-25 重庆医科大学 用于序列特异性蛋白酶检测的分子开关

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US4458066A (en) 1980-02-29 1984-07-03 University Patents, Inc. Process for preparing polynucleotides
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US6890745B1 (en) * 2000-07-19 2005-05-10 Chemicon International, Inc. Protease specific cleavable luciferases and methods of use thereof
WO2002008766A1 (fr) 2000-07-26 2002-01-31 Japan Science And Technology Corporation Sonde permettant d'analyser une interaction protéine/protéine et méthode d'analyse utilisant cette sonde
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2015530114A (ja) * 2012-09-26 2015-10-15 プロメガ コーポレイションＰｒｏｍｅｇａ Ｃｏｒｐｏｒａｔｉｏｎ リアルタイムモニタリング

Also Published As

Publication number Publication date
EP2154159A4 (en) 2012-03-21
US8658777B2 (en) 2014-02-25
EP2154159A1 (en) 2010-02-17
JPWO2008140060A1 (ja) 2010-08-05
US20100297620A1 (en) 2010-11-25
JP4491516B2 (ja) 2010-06-30

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