Substrate specificity of a protein kinases

Details Substrate specificity of a protein kinases Substrate specificity of a protein kinases

1995-01-06

1999-12-21

Celsa, Bennett

Chemistry: molecular biology and microbiology

Measuring or testing process involving enzymes or...

Involving antigen-antibody binding, specific binding protein...

435331, 436 86, 436518, 530328, 530329, 530330, 530331, 514 15, 514 16, 514 17, 514 18, G01N 3353, C07K 700

Patent

active

060047577

ABSTRACT:
The invention provides a method for determining an amino acid sequence motif for a phosphorylation site of a protein kinase. In the method of the invention, a protein kinase is contacted with an oriented degenerate peptide library, peptides within the library which are substrates for the kinase are converted to phosphopeptides and the phosphopeptides are separated from non-phosphorylated peptides. The isolated phosphopeptides are sequenced and an amino acid sequence motif for the phosphorylation site is determined based upon the relative abundance of different amino acids residues at each degenerate position. The invention also provides peptide substrates for protein kinase A, cell cycle control kinases (including cyclin B/p33.sup.cdc2 and cyclin A/p33.sup.CDK2), src family kinases (including pp60.sup.c-src and pp60.sup.v-src), EGF receptor, p92.sup.c-fps/fes, lck, c-abl, PDGF receptor, FGF receptor, insulin receptor, casein kinase II, NIMA kinase, phosphorylase kinase, Cam kinase II and Erk1 based upon amino acid sequence motifs for the phosphorylation sites of these kinases.

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