Characterization of kinase target phosphorylation consensus motifs using peptide SPOT arrays.
The human proteome is known to contain >500 protein kinases, which regulate almost all facets of cellular biology by the post-translational attachment of a phosphate moiety to serine, threonine, or tyrosine residues within a substrate protein. Most protein kinases remain poorly characterized and, as a result, current studies are directed toward defining their target substrates experimentally to gain a comprehensive view of the signaling proteins and pathways modulated by these kinases. Herein, we describe a rapid and convenient method for elucidating the consensus substrate motif for phosphorylation by a protein kinase using peptide SPOT arrays that are custom-synthesized on a cellulose membrane support. The definition of the target consensus motif provides an important starting point for the identification of physiologically relevant kinase substrates.
Leung
Genie C
GC
Therapeutics Group, The Campbell Family Institute for Breast Cancer Research, Toronto, ON, Canada.
Murphy
James M
JM
Briant
Doug
D
Sicheri
Frank
F
D020816:Amino Acid Motifs; D000818:Animals; D057075:Enzyme Assays / Q000379:methods; D006801:Humans; D008954:Models, Biological; D010449:Peptide Mapping / Q000379:methods; D010455:Peptides / Q000032:analysis / Q000138:chemical synthesis / Q000378:metabolism; D010766:Phosphorylation; D040081:Protein Array Analysis / Q000379:methods*; D011494:Protein Kinases / Q000378:metabolism*; D013379:Substrate Specificity
D010455:Peptides; D011494:Protein Kinases