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
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WO
WIPO (PCT)
Prior art keywords
protease
luc
luciferase
activated
linked
Prior art date
Application number
PCT/JP2008/058669
Other languages
English (en)
French (fr)
Inventor
Yoshio Umezawa
Takeaki Ozawa
Akira Kanno
Original Assignee
The University Of Tokyo
Inter-University Research Institute Corporation National Institutes 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 The University Of Tokyo, Inter-University Research Institute Corporation National Institutes Of Natural Sciences filed Critical The University Of Tokyo
Priority to US12/451,341 priority Critical patent/US8658777B2/en
Priority to JP2008548903A priority patent/JP4491516B2/ja
Priority to EP08752552A priority patent/EP2154159A4/en
Publication of WO2008140060A1 publication Critical patent/WO2008140060A1/ja

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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 WO2008140060A1 (ja)

Priority Applications (3)

Application Number Priority Date Filing Date Title
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
EP08752552A EP2154159A4 (en) 2007-05-09 2008-05-09 ACTIVATED PROTEASE INDICATOR

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 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 (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP1673459B1 (en) 2003-10-10 2012-06-27 Promega Corporation Luciferase biosensor
US9359635B2 (en) 2006-04-03 2016-06-07 Promega Corporation Permuted and nonpermuted luciferase biosensors
ATE542903T1 (de) 2008-05-19 2012-02-15 Promega Corp Luciferase-biosensoren für camp
JP5961163B2 (ja) 2010-05-11 2016-08-02 プロメガ コーポレイションＰｒｏｍｅｇａ Ｃｏｒｐｏｒａｔｉｏｎ 増強された検出特性を有する変異プロテアーゼバイオセンサー
US9290794B2 (en) * 2010-05-11 2016-03-22 Promega Corporation Mutant protease biosensors with enhanced detection characteristics
WO2013071237A1 (en) * 2011-11-11 2013-05-16 Promega Corporation Mutant protease biosensors with enhanced detection characteristics
JP2015509372A (ja) * 2012-03-07 2015-03-30 メルツ ファルマ ゲーエムベーハー ウント コンパニー カーゲーアーアー 修飾ルシフェラーゼに基づいて神経毒活性を測定するための手段および方法
WO2016017039A1 (ja) * 2014-07-31 2016-02-04 株式会社トランスジェニック 炎症レポーターシステム
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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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
EP2154159A1 (en) 2010-02-17
JP4491516B2 (ja) 2010-06-30
US8658777B2 (en) 2014-02-25
US20100297620A1 (en) 2010-11-25
JPWO2008140060A1 (ja) 2010-08-05
EP2154159A4 (en) 2012-03-21

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