WO2001075142A3 - Protein kinase assay - Google Patents

Protein kinase assay Download PDF

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Publication number
WO2001075142A3
WO2001075142A3 PCT/GB2001/001367 GB0101367W WO0175142A3 WO 2001075142 A3 WO2001075142 A3 WO 2001075142A3 GB 0101367 W GB0101367 W GB 0101367W WO 0175142 A3 WO0175142 A3 WO 0175142A3
Authority
WO
WIPO (PCT)
Prior art keywords
moiety
donor
protein
sample
fluorescent protein
Prior art date
Application number
PCT/GB2001/001367
Other languages
French (fr)
Other versions
WO2001075142A2 (en
Inventor
Jeremy Myles Tavare
Gavin Iain Welsh
Original Assignee
Univ Bristol
Jeremy Myles Tavare
Gavin Iain Welsh
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 Univ Bristol, Jeremy Myles Tavare, Gavin Iain Welsh filed Critical Univ Bristol
Priority to AU44316/01A priority Critical patent/AU4431601A/en
Publication of WO2001075142A2 publication Critical patent/WO2001075142A2/en
Publication of WO2001075142A3 publication Critical patent/WO2001075142A3/en

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Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12QMEASURING 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
    • C12Q1/00Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/48Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase
    • C12Q1/485Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving transferase involving kinase

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  • 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)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Molecular Biology (AREA)
  • Biotechnology (AREA)
  • Biophysics (AREA)
  • Analytical Chemistry (AREA)
  • Immunology (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)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)

Abstract

An assay for the activity of a kinase enzyme contained in a sample, which comprises: i) contacting the sample with a pair of protein constructs, one of which comprises a donor fluorescent protein and the other of which comprises an acceptor fluorescent protein moiety, the donor protein moiety and the acceptor protein moiety being selected such that, when the said fluorescent protein moieties are in sufficiently close proximity, they exhibit resonance energy transfer when the donor moiety is excited, wherein a first one of said constructs comprises a target peptide moiety having an amino acid sequence which is a target for phosphorylation by the kinase enzyme, and the other (the second) of which comprises a binding protein moiety which either: a) selectively binds to the target peptide moiety of the first construct only in its phosphorylated form; or b) selectively binds to the target peptide moiety of the first construct only in its non-phosphorylated form; to form a complex in which the donor and acceptor protein moieties are brought into sufficiently close proximity that resonance energy transfer is exhibited upon excitation of the donor moiety; ii) exciting the donor fluorescent protein moiety; iii) determining the degree of fluorescence resonance energy transfer in the sample.
PCT/GB2001/001367 2000-03-31 2001-03-27 Protein kinase assay WO2001075142A2 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
AU44316/01A AU4431601A (en) 2000-03-31 2001-03-27 Protein kinase assay

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB0007915.2 2000-03-31
GB0007915A GB0007915D0 (en) 2000-03-31 2000-03-31 Protein kinase assay

Publications (2)

Publication Number Publication Date
WO2001075142A2 WO2001075142A2 (en) 2001-10-11
WO2001075142A3 true WO2001075142A3 (en) 2002-05-16

Family

ID=9888902

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB2001/001367 WO2001075142A2 (en) 2000-03-31 2001-03-27 Protein kinase assay

Country Status (3)

Country Link
AU (1) AU4431601A (en)
GB (1) GB0007915D0 (en)
WO (1) WO2001075142A2 (en)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003087394A1 (en) * 2002-04-15 2003-10-23 Glaxo Group Limited A method of identifying a modudator for a serine/theronine kinase
US20070264678A1 (en) 2005-10-28 2007-11-15 Invitrogen Corporation Kinase and ubiquination assays
CN104204799A (en) * 2012-02-06 2014-12-10 珀金埃尔默生物信号股份有限公司 Dual-acceptor time-resolved-fret
US10914728B2 (en) 2016-10-24 2021-02-09 Novo Nordisk A/S Bioassay for insulin formulations

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998018956A1 (en) * 1996-10-28 1998-05-07 Panvera Corporation Kinase activity measurement using fluorescence polarization
WO1999029894A1 (en) * 1997-12-05 1999-06-17 Pharmacia & Upjohn Company Fluorescence-based high throughput screening assays for protein kinases and phosphatases
WO2000043780A1 (en) * 1999-01-19 2000-07-27 Imperial Cancer Research Technology Limited Methods for detecting changes to cellular components
WO2000050896A1 (en) * 1999-02-25 2000-08-31 Cyclacel Limited Compositions and methods for monitoring the modification of engineered binding partners
WO2000075167A2 (en) * 1999-06-09 2000-12-14 Ljl Biosystems, Inc. Phosphorylation assays

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO1998018956A1 (en) * 1996-10-28 1998-05-07 Panvera Corporation Kinase activity measurement using fluorescence polarization
WO1999029894A1 (en) * 1997-12-05 1999-06-17 Pharmacia & Upjohn Company Fluorescence-based high throughput screening assays for protein kinases and phosphatases
WO2000043780A1 (en) * 1999-01-19 2000-07-27 Imperial Cancer Research Technology Limited Methods for detecting changes to cellular components
WO2000050896A1 (en) * 1999-02-25 2000-08-31 Cyclacel Limited Compositions and methods for monitoring the modification of engineered binding partners
WO2000075167A2 (en) * 1999-06-09 2000-12-14 Ljl Biosystems, Inc. Phosphorylation assays

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
NG TONY ET AL: "Imaging protein kinase Calpha activation in cells.", SCIENCE (WASHINGTON D C), vol. 283, no. 5410, 26 March 1999 (1999-03-26), pages 2085 - 2089, XP002191053, ISSN: 0036-8075 *
WOUTERS F S ET AL: "FLUORESCENCE LIFETIME IMAGING OF RECEPTOR TYROSINE KINASE ACTIVITY IN CELLS", CURRENT BIOLOGY, CURRENT SCIENCE,, GB, vol. 9, no. 19, 27 September 1999 (1999-09-27), pages 1127 - 1130, XP001036554, ISSN: 0960-9822 *

Also Published As

Publication number Publication date
GB0007915D0 (en) 2000-05-17
WO2001075142A2 (en) 2001-10-11
AU4431601A (en) 2001-10-15

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