Screening phage-displayed random peptide libraries for SH3 ligands

A. B. Sparks, N. B. Adey, Lawrence Quilliam, J. M. Thorn, B. K. Kay

Research output: Contribution to journalArticle

26 Citations (Scopus)

Abstract

Using GST-Src SH3 fusion protein to screen the phage-displayed RPLs T9 and T12, we have isolated biologically active peptide ligands for, and characterized the binding specificity of, the Src SH3 domain. Peptides displayed by Src SH3 affinity-purified phage contain similar proline-rich regions that yield a consensus motif of RPLPPLP (Fig. 3); this motif is similar to a region of p85 phosphatidylinositol 3'-kinase known to interact with Src SH3. Phage expressing peptides related to the RPLPPLP motif bind GST-SrcSH3, but not GST-Abl SH3 or GST alone (Fig. 4). Similarly, synthetic peptides containing the RPLPPLP motif compete Src SH3-binding proteins, but not Abl SH3- or PLCγ SH3-binding proteins, from cell lysates. Finally, RPLPPLP-related peptides are able to accelerate progesterone-induced maturation of Xenopus laevis oocytes, a similar acceleration is observed in oocytes treated with activated, but not normal, Xenopus Src. These results suggest that screening nonbiased peptide libraries may represent a general means of isolating peptide ligands for, and characterizing the binding specificities of, other domains thought to mediate protein-protein interactions, including other SH3 domains, armadillo repeats, and PH domains.

Original languageEnglish (US)
Pages (from-to)498-509
Number of pages12
JournalMethods in Enzymology
Volume255
DOIs
StatePublished - 1995
Externally publishedYes

Fingerprint

Peptide Library
Bacteriophages
Screening
Ligands
Peptides
Src peptide
src Homology Domains
Oocytes
Carrier Proteins
Phosphatidylinositol 3-Kinase
Armadillos
Proteins
Xenopus laevis
Programmable logic controllers
Xenopus
Proline
Progesterone
Fusion reactions

ASJC Scopus subject areas

  • Biochemistry

Cite this

Screening phage-displayed random peptide libraries for SH3 ligands. / Sparks, A. B.; Adey, N. B.; Quilliam, Lawrence; Thorn, J. M.; Kay, B. K.

In: Methods in Enzymology, Vol. 255, 1995, p. 498-509.

Research output: Contribution to journalArticle

Sparks, A. B. ; Adey, N. B. ; Quilliam, Lawrence ; Thorn, J. M. ; Kay, B. K. / Screening phage-displayed random peptide libraries for SH3 ligands. In: Methods in Enzymology. 1995 ; Vol. 255. pp. 498-509.
@article{6723069fe6dd48319088d151748692a7,
title = "Screening phage-displayed random peptide libraries for SH3 ligands",
abstract = "Using GST-Src SH3 fusion protein to screen the phage-displayed RPLs T9 and T12, we have isolated biologically active peptide ligands for, and characterized the binding specificity of, the Src SH3 domain. Peptides displayed by Src SH3 affinity-purified phage contain similar proline-rich regions that yield a consensus motif of RPLPPLP (Fig. 3); this motif is similar to a region of p85 phosphatidylinositol 3'-kinase known to interact with Src SH3. Phage expressing peptides related to the RPLPPLP motif bind GST-SrcSH3, but not GST-Abl SH3 or GST alone (Fig. 4). Similarly, synthetic peptides containing the RPLPPLP motif compete Src SH3-binding proteins, but not Abl SH3- or PLCγ SH3-binding proteins, from cell lysates. Finally, RPLPPLP-related peptides are able to accelerate progesterone-induced maturation of Xenopus laevis oocytes, a similar acceleration is observed in oocytes treated with activated, but not normal, Xenopus Src. These results suggest that screening nonbiased peptide libraries may represent a general means of isolating peptide ligands for, and characterizing the binding specificities of, other domains thought to mediate protein-protein interactions, including other SH3 domains, armadillo repeats, and PH domains.",
author = "Sparks, {A. B.} and Adey, {N. B.} and Lawrence Quilliam and Thorn, {J. M.} and Kay, {B. K.}",
year = "1995",
doi = "10.1016/S0076-6879(95)55052-6",
language = "English (US)",
volume = "255",
pages = "498--509",
journal = "ImmunoMethods",
issn = "1046-2023",
publisher = "Academic Press Inc.",

}

TY - JOUR

T1 - Screening phage-displayed random peptide libraries for SH3 ligands

AU - Sparks, A. B.

AU - Adey, N. B.

AU - Quilliam, Lawrence

AU - Thorn, J. M.

AU - Kay, B. K.

PY - 1995

Y1 - 1995

N2 - Using GST-Src SH3 fusion protein to screen the phage-displayed RPLs T9 and T12, we have isolated biologically active peptide ligands for, and characterized the binding specificity of, the Src SH3 domain. Peptides displayed by Src SH3 affinity-purified phage contain similar proline-rich regions that yield a consensus motif of RPLPPLP (Fig. 3); this motif is similar to a region of p85 phosphatidylinositol 3'-kinase known to interact with Src SH3. Phage expressing peptides related to the RPLPPLP motif bind GST-SrcSH3, but not GST-Abl SH3 or GST alone (Fig. 4). Similarly, synthetic peptides containing the RPLPPLP motif compete Src SH3-binding proteins, but not Abl SH3- or PLCγ SH3-binding proteins, from cell lysates. Finally, RPLPPLP-related peptides are able to accelerate progesterone-induced maturation of Xenopus laevis oocytes, a similar acceleration is observed in oocytes treated with activated, but not normal, Xenopus Src. These results suggest that screening nonbiased peptide libraries may represent a general means of isolating peptide ligands for, and characterizing the binding specificities of, other domains thought to mediate protein-protein interactions, including other SH3 domains, armadillo repeats, and PH domains.

AB - Using GST-Src SH3 fusion protein to screen the phage-displayed RPLs T9 and T12, we have isolated biologically active peptide ligands for, and characterized the binding specificity of, the Src SH3 domain. Peptides displayed by Src SH3 affinity-purified phage contain similar proline-rich regions that yield a consensus motif of RPLPPLP (Fig. 3); this motif is similar to a region of p85 phosphatidylinositol 3'-kinase known to interact with Src SH3. Phage expressing peptides related to the RPLPPLP motif bind GST-SrcSH3, but not GST-Abl SH3 or GST alone (Fig. 4). Similarly, synthetic peptides containing the RPLPPLP motif compete Src SH3-binding proteins, but not Abl SH3- or PLCγ SH3-binding proteins, from cell lysates. Finally, RPLPPLP-related peptides are able to accelerate progesterone-induced maturation of Xenopus laevis oocytes, a similar acceleration is observed in oocytes treated with activated, but not normal, Xenopus Src. These results suggest that screening nonbiased peptide libraries may represent a general means of isolating peptide ligands for, and characterizing the binding specificities of, other domains thought to mediate protein-protein interactions, including other SH3 domains, armadillo repeats, and PH domains.

UR - http://www.scopus.com/inward/record.url?scp=0029129806&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0029129806&partnerID=8YFLogxK

U2 - 10.1016/S0076-6879(95)55052-6

DO - 10.1016/S0076-6879(95)55052-6

M3 - Article

C2 - 8524137

AN - SCOPUS:0029129806

VL - 255

SP - 498

EP - 509

JO - ImmunoMethods

JF - ImmunoMethods

SN - 1046-2023

ER -