Development and characterization of potent and specific peptide inhibitors of p60c-src protein tyrosine kinase using pseudosubstrate-based inhibitor design approach

J. R. Kamath, Ruiwu Liu, A. M. Enstrom, Q. Lou, Kit Lam

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

The cytoplasmic protein p60c-src, an ubiquitous non-receptor protein tyrosine kinase (PTK) is a potential anticancer target as it is over-expressed and/or constitutively active in several cancer types. In addition, the phenotype of c-src knock-out mice is consistent with osteopetrosis, which suggests that inhibitors against this enzyme may also be therapeutic for osteoporosis. Using a known peptide substrate for c-src, MIYKYYF, as a template, we have developed a series of pseudosubstrate-based peptide inhibitors. Structure-activity relationship studies have been performed on one of these inhibitors, CIYKYYF. In a kinase assay using YIYGSFK as the substrate, CIYKYY has been demonstrated to inhibit p60c-src, with an IC50 of 0.6 μM. Further truncation has led to the determination that even the smaller peptide, CIYK, is a moderately potent inhibitor with IC50 of 15 μM. Some improvement in inhibitory potency (IC 50 = 11.8 μM) has been observed with the replacement of Tyr 3 in CIYK with β-phenylalanine (β-Phe). The tetrapeptide CI(β-Phe)K will be used as a lead compound for future development of peptidomimetics and small molecule inhibitors that have the capacity to penetrate the plasma membrane of intact cells.

Original languageEnglish (US)
Pages (from-to)260-268
Number of pages9
JournalJournal of Peptide Research
Volume62
Issue number6
DOIs
StatePublished - Dec 2003

Fingerprint

src-Family Kinases
Peptides
Inhibitory Concentration 50
Osteopetrosis
Peptidomimetics
Lead compounds
Enzyme Inhibitors
Substrates
Cell membranes
Structure-Activity Relationship
Phenylalanine
Knockout Mice
Protein-Tyrosine Kinases
Osteoporosis
Assays
Phosphotransferases
Cell Membrane
Phenotype
Molecules
Enzymes

Keywords

  • p60 protein tyrosine kinase
  • Protein kinase assay with thin layer chromatography
  • Pseudosubstrate-based peptide inhibitors
  • Structure-activity relationship study

ASJC Scopus subject areas

  • Biochemistry
  • Endocrinology

Cite this

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abstract = "The cytoplasmic protein p60c-src, an ubiquitous non-receptor protein tyrosine kinase (PTK) is a potential anticancer target as it is over-expressed and/or constitutively active in several cancer types. In addition, the phenotype of c-src knock-out mice is consistent with osteopetrosis, which suggests that inhibitors against this enzyme may also be therapeutic for osteoporosis. Using a known peptide substrate for c-src, MIYKYYF, as a template, we have developed a series of pseudosubstrate-based peptide inhibitors. Structure-activity relationship studies have been performed on one of these inhibitors, CIYKYYF. In a kinase assay using YIYGSFK as the substrate, CIYKYY has been demonstrated to inhibit p60c-src, with an IC50 of 0.6 μM. Further truncation has led to the determination that even the smaller peptide, CIYK, is a moderately potent inhibitor with IC50 of 15 μM. Some improvement in inhibitory potency (IC 50 = 11.8 μM) has been observed with the replacement of Tyr 3 in CIYK with β-phenylalanine (β-Phe). The tetrapeptide CI(β-Phe)K will be used as a lead compound for future development of peptidomimetics and small molecule inhibitors that have the capacity to penetrate the plasma membrane of intact cells.",
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AU - Liu, Ruiwu

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AU - Lou, Q.

AU - Lam, Kit

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