Identification and Characterization of Src SH3 Ligands from Phage-displayed Random Peptide Libraries

Andrew B. Sparks, Lawrence Quilliam, Judith M. Thorn, Channing J. Der, Brian K. Kay

Research output: Contribution to journalArticle

147 Citations (Scopus)

Abstract

We have used the Src homology 3 (SH3) domain to screen two phage-displayed random peptide libraries, each containing 2 × 108 unique members, and have identified a series of high affinity peptide ligands. The peptides possess similar proline-rich regions, which yield a consensus Src SH3-binding motif of RPLPPLP. We have confirmed this motif by screening a phage-displayed peptide library biased for SH3 ligands and identifying the same consensus sequence. Binding studies using synthetic peptides suggest that the RPLPPLP motif is important for SH3 binding and confers specificity for the Src SH3 domain, and that residues which flank the motif may also contribute to binding. Peptides that contain the RPLPPLP motif compete Src, but not Abl or phospholipase Cγ, SH3 interactions with SH3-binding proteins from cell lysates (IC50 = 1-5 μM). Furthermore, RPLPPLP-related peptides are able to accelerate progesterone-induced maturation of Xenopus laevis oocytes. A similar acceleration has been observed in oocytes treated with activated, but not normal, Xenopus Src, suggesting the possibility that the peptides are able to antagonize the negative regulation of Src activity by Src SH3 in vivo.

Original languageEnglish (US)
Pages (from-to)23853-23856
Number of pages4
JournalJournal of Biological Chemistry
Volume269
Issue number39
StatePublished - Sep 30 1994
Externally publishedYes

Fingerprint

Peptide Library
Bacteriophages
Ligands
Peptides
src Homology Domains
Oocytes
Consensus Sequence
Xenopus laevis
Type C Phospholipases
Xenopus
Proline
Inhibitory Concentration 50
Progesterone
Carrier Proteins
Screening

ASJC Scopus subject areas

  • Biochemistry

Cite this

Identification and Characterization of Src SH3 Ligands from Phage-displayed Random Peptide Libraries. / Sparks, Andrew B.; Quilliam, Lawrence; Thorn, Judith M.; Der, Channing J.; Kay, Brian K.

In: Journal of Biological Chemistry, Vol. 269, No. 39, 30.09.1994, p. 23853-23856.

Research output: Contribution to journalArticle

Sparks, Andrew B. ; Quilliam, Lawrence ; Thorn, Judith M. ; Der, Channing J. ; Kay, Brian K. / Identification and Characterization of Src SH3 Ligands from Phage-displayed Random Peptide Libraries. In: Journal of Biological Chemistry. 1994 ; Vol. 269, No. 39. pp. 23853-23856.
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