Journal article
Detection of Src homology 3-binding proteins, including paxillin, in normal and v-Src-transformed Balb/c 3T3 cells
The Journal of biological chemistry, v 268(20), pp 14956-14963
15 Jul 1993
PMID: 8325872
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
The Src homology 3 (SH3) domain, located in the amino-terminal, noncatalytic half of pp60 super(src), is highly conserved among members of the Src family of tyrosine kinases. SH3 domains have also been identified in a variety of proteins otherwise unrelated to protein-tyrosine kinases. The presence of SH3 domains in proteins with diverse functions suggests this domain may be important for directing protein-protein interactions necessary for protein function or cellular localization. To explore possible interactions between the SH3 domain and cellular proteins, we have established conditions for the isolation of proteins that bind in solution to the Src SH3 domain. A 67-amino acid fragment of c-Src containing either the entire glutathione S-transferase-SH3 domain (GST-SH3) or the SH3 domain from the neuronal form of c-Src (GST-SH3 super(+)) was expressed as a glutathione S-transferase fusion protein. The GST fusion proteins were incubated with lysates from ( super(35)S)methionine-labeled Balb/c 3T3 cells or v-Src-transformed Balb/c 3T3 cells. We found that GST-SH3, but not wild-type GST, specifically interacted with multiple cellular proteins, whereas GST-SH3 super(+) only weakly associated with a small subset of these proteins. The majority of the SH3-binding proteins were found in particulate and detergent-insoluble cell fractions. Anti-phosphotyrosine immunoblots of the SH3-binding proteins revealed that several of the SH3-binding proteins are phosphorylated on tyrosine in v-Src-transformed cells. In addition, a number of the SH3-binding proteins were phosphorylated on serine and/or threonine in in vitro kinase assays, suggesting that one or more of the SH3-binding proteins has kinase activity. We identified paxillin, a vinculin-binding protein, as one of the Src SH3-binding proteins.
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Details
- Title
- Detection of Src homology 3-binding proteins, including paxillin, in normal and v-Src-transformed Balb/c 3T3 cells
- Creators
- Zhigang WengJ A TaylorC E TurnerJ BruggeC Seidel-Dugan
- Publication Details
- The Journal of biological chemistry, v 268(20), pp 14956-14963
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Environmental and Occupational Health
- Web of Science ID
- WOS:A1993LL75900064
- Scopus ID
- 2-s2.0-0027293670
- Other Identifier
- 991021888920604721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Biochemistry & Molecular Biology