WO2005114198A3 - Protein phosphorylation assay - Google Patents

Protein phosphorylation assay Download PDF

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Publication number
WO2005114198A3
WO2005114198A3 PCT/US2005/017711 US2005017711W WO2005114198A3 WO 2005114198 A3 WO2005114198 A3 WO 2005114198A3 US 2005017711 W US2005017711 W US 2005017711W WO 2005114198 A3 WO2005114198 A3 WO 2005114198A3
Authority
WO
WIPO (PCT)
Prior art keywords
substrate
labeled
phosphatases
enzymes
bind
Prior art date
Application number
PCT/US2005/017711
Other languages
French (fr)
Other versions
WO2005114198A2 (en
Inventor
Douglas S Auld
Original Assignee
Pharmacopeia Inc
Douglas S Auld
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 Pharmacopeia Inc, Douglas S Auld filed Critical Pharmacopeia Inc
Publication of WO2005114198A2 publication Critical patent/WO2005114198A2/en
Publication of WO2005114198A3 publication Critical patent/WO2005114198A3/en

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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
    • C12Q1/42—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving hydrolase involving phosphatase
    • 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
    • C12Q1/485—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase involving kinase
    • 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

Landscapes

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

Abstract

The present invention provides methods for assaying the activity of protein kinases or phosphatases that involve reacting the enzymes with substrate, at least a portion of which is fluorescently labeled; allowing phosphorylated substrate to bind to a solid support coated with a phosphate-binding material; and measuring the fluorescence produced by bound, labeled substrate. The invention also provides methods for identifying substances that modulate the activity of protein kinases and phosphatases that comprise reacting the enzymes with a substrate, at least a portion of which is fluorescently labeled, in the presence and absence of test compounds, allowing phosphorylated substrate to bind to a solid support coated with a phosphate-binding material, measuring the fluorescence produced by bound, labeled substrate, and comparing the fluorescence from reactions carried out in the presence and absence of test compounds.
PCT/US2005/017711 2004-05-20 2005-05-19 Protein phosphorylation assay WO2005114198A2 (en)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US57274104P 2004-05-20 2004-05-20
US60/572,741 2004-05-20

Publications (2)

Publication Number Publication Date
WO2005114198A2 WO2005114198A2 (en) 2005-12-01
WO2005114198A3 true WO2005114198A3 (en) 2006-05-26

Family

ID=35428998

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/US2005/017711 WO2005114198A2 (en) 2004-05-20 2005-05-19 Protein phosphorylation assay

Country Status (1)

Country Link
WO (1) WO2005114198A2 (en)

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
FI20070171A0 (en) * 2007-02-28 2007-02-28 Hidex Oy Bioassay and bioassay equipment
CN115057472B (en) * 2022-06-21 2023-10-27 中国医学科学院基础医学研究所 Novel fluorescence sensing system and application thereof in PTP-1B detection

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001025477A2 (en) * 1999-10-01 2001-04-12 Biofocus Discovery Limited Kinase and phosphatase assays
US6942987B2 (en) * 2002-05-15 2005-09-13 Pharmacopeia Drug Discovery, Inc. Methods for measuring kinase activity

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2001025477A2 (en) * 1999-10-01 2001-04-12 Biofocus Discovery Limited Kinase and phosphatase assays
US6942987B2 (en) * 2002-05-15 2005-09-13 Pharmacopeia Drug Discovery, Inc. Methods for measuring kinase activity

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
BEASLEY J ET AL: "Evaluation of Compound Interference in Immobilized Metal Ion Affinity-Based Fluorescence Polarization Detection with a Four Million Member Compound Collection.", ASSAY AND DRUG DEVELOPMENT TECHNOLOGIES., vol. 1, no. 3, 2003, pages 455 - 459, XP008062391 *

Also Published As

Publication number Publication date
WO2005114198A2 (en) 2005-12-01

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