PMID: 7536039Apr 5, 1995Paper

Use of synthetic peptides and copolymers to study the substrate specificity and inhibition of the protein tyrosine kinase pp60c-src

Biochimica Et Biophysica Acta
R J BuddeJ S McMurray

Abstract

The ability of synthetic peptides and polypeptides to act as substrates and/or inhibitors of pp60c-src was examined. The random copolymer, poly(K4Y) had a threefold lower specificity than poly(E4Y). Peptides containing lysine vs. glutamate were also found to have a lower substrate specificity (Vmax:Km ratio). In order to assess the substrate specificity of acidic peptides, an assay protocol using DEAE-membranes was developed. Peptides containing a (YXE)5YXD motif (X = G, A, V, P, or norvaline) were tested as inhibitors and substrates of pp60c-src. The glycine-containing peptide was the best substrate having a specificity 16,000-fold higher than 5Val-angiotensin II, the most commonly used peptide substrate. Most of the peptides, except for the proline containing peptide, had Ki values of 20-100 microM. In a series of (XGE)5XGD peptides, where X = Y or F, tyrosine at position 10 was found to be the preferred site for accepting a phosphate. Analogs in which the glycine was replaced with alanine indicated that loss of flexibility around position 10 was detrimental to substrate specificity. Results suggest that conformational requirements of the peptides tested was important and substrate specificity was a more sensitive parameter t...Continue Reading

References

Feb 1, 1992·Analytical Biochemistry·R J BuddeD A Tinker
Jul 27, 1990·Science·J K Scott, G P Smith
Jan 1, 1991·Methods in Enzymology·D O MorganH E Varmus
Mar 1, 1990·BioEssays : News and Reviews in Molecular, Cellular and Developmental Biology·R K RalphP Schofield
Feb 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·T PatschinskyB M Sefton
Jan 1, 1984·Advances in Enzyme Regulation·J E Casnellie, E G Krebs
Sep 1, 1993·International Journal of Peptide and Protein Research·J S McMurrayD F Dyckes
Feb 1, 1994·International Journal of Peptide and Protein Research·N U ObeyesekereJ S McMurray

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Citations

Jun 12, 1999·Journal of Immunological Methods·G R Nakayama, Z Parandoosh
Mar 29, 2011·TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik·A C CrawfordM G Francki
May 15, 2002·Bioscience, Biotechnology, and Biochemistry·Hiroyuki NagataTatsuya Tamaoki
Dec 12, 1996·Biochimica Et Biophysica Acta·L A Pinna, M Ruzzene

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