The functions of Src family kinases are tightly regulated through Src homology (SH) domain-mediated protein-protein interactions. We previously reported the biophysical characteristics of the apoptosis-linked gene 2-interacting protein X (Alix) in complex with the haemopoietic cell kinase (Hck) SH3 domain. In the current study, we have combined ITC, NMR, SAXS and molecular modeling to determine a 3D model of the complex. We demonstrate that Hck SH3 recognizes an extended linear proline-rich region of Alix. This particular binding mode enables Hck SH3 to sense a specific non-canonical residue situated in the SH3 RT-loop of the kinase. The resulting model helps clarify the mechanistic insights of Alix-Hck interaction. Structured summary of protein interactions: Hck physically interacts with SAM68 by two hybrid (View interaction) FynR96I physically interacts with Alix by two hybrid (View interaction) Hck binds to Alix by pull down (View interaction) Fyn physically interacts with SAM68 by two hybrid (View interaction) Hck and Alix bind by nuclear magnetic resonance (View interaction) FynR96I and Alix bind by isothermal titration calorimetry (View Interaction: 1, 2) FynR96I and Alix bind by nuclear magnetic resonance (View interaction) FynR96I binds to Alix by pull down (View interaction) Hck physically interacts with Alix by two hybrid (View interaction) FynR96I and Alix bind by x ray scattering (View interaction) Hck physically interacts with NEF by two hybrid (View interaction) FynR96I physically interacts with NEF by two hybrid (View interaction).
|Original language||English (US)|
|Number of pages||6|
|State||Published - Jun 21 2012|
Bibliographical noteFunding Information:
This work was supported by the Centre National de la Recherche Scientifique (CNRS); the Institut National de la Santé et de la Recherche Médicale (INSERM); the Agence Nationale de Recherche sur le SIDA et les hépatites virales (ANRS); the Ministry of Science and Technology, China [Grant Nos. 2007CB914304; 2006AA02A313]; the National Natural Science Foundation of China (NSFC) [Grant Nos. 30800181; 30625011]; the Research Foundation Flanders (FWO); the concerted action programme of the Katholieke Universiteit Leuven; and the National Institutes of Health (MD Anderson’s Cancer Center Support) [Grant No. CA016672]. S.O. and A.L. were fellows of the ANRS, and X.S. was a fellow of the ‘Ambassade de France en Chine’.
- Apoptosis-linked gene 2-interacting protein X (Alix)
- Protein-protein interaction
- Src family kinase (SFK)
- Src homology 3 domain (SH3 domain)
ASJC Scopus subject areas
- Structural Biology
- Molecular Biology
- Cell Biology