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US 6,670,144 B1Granted
Compositions and methods for monitoring the phosphorylation of natural binding partners
AssigneeCyclacel Limited
InventorsRoger K. Craig, John Colyer
Life of this patent
Estimating the term…
Abstract
This invention relates to methods and compositions for monitoring the interaction of binding partners as a function of the addition or subtraction of a phosphate group to or from one of the binding partners by a protein kinase or phosphatase.
Claims6
A method of screening for a candidate modulator of enzymatic activity of a kinase or a phosphatase, the method comprising
- a) contacting an isolated natural binding domain, a binding partner therefor and an enzyme with a candidate modulator of said kinase or phosphatase, wherein said natural binding domain includes a site for post-translational phosphorylation and binds said binding partner in a manner that is dependent upon phosphorylation or dephosphorylation of said site by said kinase or phosphatase and wherein at least one of said isolated natural binding domain and said binding partner comprises a detectable label;
- b) detecting the binding or dissociation of said isolated natural binding domain to/from said binding partner;
- and
- c) monitoring the binding of said isolated natural binding domain to said binding partner, wherein binding or dissociation.of said isolated natural binding domain and said binding partner as a result of said contacting is indicative of modulation of enzymatic activity by said candidate modulator of said kinase or phosphatase.
1 more element
A method of screening for a candidate modulator of enzymatic activity of a kinase or a phosphatase, the method comprising
- a) contacting a cell-free assay system comprising an isolated natural binding domain and a binding partner therefor with a candidate modulator of enzymatic activity of a said kinase or phosphatase;
- b) detecting the binding or dissociation of said isolated natural binding domain to/from said binding partner;
- and
- c) monitoring binding of said isolated natural binding domain and said binding partner therefor in said assay system, wherein said isolated natural binding domain includes a site for post-translational phosphorylation and binds said binding partner in a manner that is dependent upon phosphorylation or dephosphorylation of said site by a said kinase or phosphatase in said assay system, wherein at least one of said isolated natural binding domain and said binding partner comprises a detectable label, and wherein binding or dissociation of said isolated natural binding domain and said binding partner as a result of said contacting is indicative of modulation of enzymatic activity by said candidate modulator of a said kinase or phosphatase.
1 more element
Dependencies are inferred from the claim text.