A yeast two‐hybrid study of human p97/Gab2 interactions with its SH2 domain‐containing binding partners

FEBS Letters

Camonis, Jacques¹  Gesbert, Franck²  Arnaud, Mary²  Crouin, Catherine²  Bertoglio, Jacques² 

[1] Inserm Unit 528, Institut Curie, 26 rue d'Ulm, 75248 Paris, France;Inserm Unit 461, Faculté de Pharmacie Paris-XI, 5 rue Jean-Baptiste Clément, 92296 Châtenay-Malabry, France

关键词: Src homology 2 domain;    Phosphoinositide-3-kinase;    SHP-2;    CrkL;    Gab2;    Yeast two-hybrid;    SH2;    Src homology 2 domain;    PI3K;    phosphoinositide-3-kinase;    IRS;    insulin receptor substrate;    X-gal;    5-bromo-4-chloro-3-indolyl-β-D-galactopyranoside;    ONPG;    2-nitrophenyl-β-D-galactopyranoside;   

DOI  :  10.1016/S0014-5793(01)02373-0

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

p97/Gab2 is a recently characterized member of a large family of scaffold proteins that play essential roles in signal transduction. Gab2 becomes tyrosine-phosphorylated in response to a variety of growth factors and forms multimolecular complexes with SH2 domain-containing signaling molecules such as the p85-regulatory subunit of the phosphoinositide-3-kinase (p85-PI3K), the tyrosine phosphatase SHP-2 and the adapter protein CrkL. To characterize the interactions between Gab2 and its SH2-containing binding partners, we designed a modified yeast two-hybrid system in which the Lyn tyrosine kinase is expressed in a regulated manner in yeast. Using this assay, we demonstrated that p97/Gab2 specifically interacts with the SH2 domains of PI3K, SHP-2 and CrkL. Interaction with p85-PI3K is mediated by tyrosine residues Y₄₅₂, Y₄₇₆ and Y₅₈₄ of Gab2, while interaction with SHP-2 depends exclusively on tyrosine Y₆₁₄. CrkL interaction is mediated by its SH2 domain recognizing Y₂₆₆ and Y₂₉₃, despite the latter being in a non-consensus (YTFK) environment.

【 授权许可】

Unknown