Project IV: Nef ? Hck interaction (T. Smithgall, Cores C, D, and E) - Large amounts of soluble Nef for preliminary NMR evaluation have been obtained. The regulatory region of Hck (Hck 32L, containing the SH3 and SH2 domains as well as the SH2-kinase linker), has been expressed in E. coli with very high yield. When combined in an equimolar ratio, a single, stable Nef-Hck32L complex is formed. Preliminary conditions for Nef-Hck crystallization have been identified using developed protocols. - Hck32L has been triple-labeled and ongoing NMR experiments are expected to reveal chemical shift changes within the SH3 domain and elsewhere upon Nef binding.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Specialized Center (P50)
Project #
5P50GM082251-02
Application #
7671237
Study Section
Special Emphasis Panel (ZRG1)
Project Start
Project End
Budget Start
2008-08-01
Budget End
2009-07-31
Support Year
2
Fiscal Year
2008
Total Cost
$145,022
Indirect Cost
Name
University of Pittsburgh
Department
Type
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
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