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Tyrosine phosphorylation-dependent yeast two-hybrid system for the identification of the SH2 domain-binding proteins

Title
Tyrosine phosphorylation-dependent yeast two-hybrid system for the identification of the SH2 domain-binding proteins
Authors
Park D.Yun Y.
Ewha Authors
윤영대
SCOPUS Author ID
윤영대scopus
Issue Date
2001
Journal Title
Molecules and Cells
ISSN
1016-8478JCR Link
Citation
vol. 12, no. 2, pp. 244 - 249
Indexed
SCI; SCIE; SCOPUS; KCI WOS scopus
Abstract
In this paper, we established a modified yeast two-hybrid system, which is specialized for the detection of SH2 domain-binding proteins. The employment of the SH2 domain-tyrosine kinase fusion protein as bait allowed the efficient identification of SH2 domain-binding proteins. The general applicability of the system was tested using various combinations of SH2-kinase fusion bait and prey. The results indicate that the system specifically detected the previously reported in vivo interactions between the SH2 domains and their binding partners. In addition, using this system, we found the interaction between the adaptor protein, Lad, and the SH2 domain of Grb2 or PLC-γ1. The binding of Lad to Grb2 was further confirmed in mammalian cells by a co-immunoprecipitation study. The conclusion is that the established tyrosine phosphorylation-dependent yeast two-hybrid system provides a novel and efficient way to define the SH2 domain-binding molecules. ©KSMCB 2001.
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자연과학대학 > 생명과학전공 > Journal papers
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