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Identification of 3BP-1 in cDNA expression library by SH3 domain screening

Cicchetti, Piera and Baltimore, David (1995) Identification of 3BP-1 in cDNA expression library by SH3 domain screening. In: Small GTPases and Their Regulators Part B: Rho Family. Methods in Enzymology. No.256. Academic Press , San Diego, CA, pp. 140-148. ISBN 978-0-12-182157-9.

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This chapter describes a screening procedure to assay for protein–protein interactions using a biotinylated glutathione S-transferase (GST) fusion protein probe to screen cDNA expression libraries. This method is developed to allow nonradioactive probing, which has the additional advantages of a very low background signal and a low incidence of false positives. Because neither the fusion protein probe nor the protein in the expression library is denatured, this assay allows the structural components of the binding reaction to remain intact. This screening procedure offers a practical and highly efficient method of identifying protein–protein interactions. Using this procedure to detect proteins that bound to the Abl SH3 domain, five cDNA clones were isolated out of 7 million cDNA containing plaques that were screened. Of these five, three contained the identical cDNA clone termed “3BP-1,” while the other two were identical for a different clone termed “3BP-2.” On sequencing these clones, only a short stretch of approximately 40 amino acids was found where sequence similarity existed between these proteins.

Item Type:Book Section
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Baltimore, David0000-0001-8723-8190
Additional Information:© 1995 by Academic Press, Inc.
Series Name:Methods in Enzymology
Issue or Number:256
Record Number:CaltechAUTHORS:20200512-154625286
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Official Citation:Piera Cicchetti, David Baltimore, Identification of 3BP-1 in cDNA expression library by SH3 domain screening, Methods in Enzymology, Academic Press, Volume 256, 1995, Pages 140-148, ISSN 0076-6879, ISBN 9780121821579, (
Usage Policy:No commercial reproduction, distribution, display or performance rights in this work are provided.
ID Code:103160
Deposited By: Tony Diaz
Deposited On:12 May 2020 23:09
Last Modified:16 Nov 2021 18:18

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