Solution Structure of BAG Domains: BAG-1 delta N
EMSL Project ID
2002
Abstract
BAG-1 is a novel multifunctional protein first identified as a molecule that binds to the anti-apoptotic protein Bcl-2. With the discovery that BAG-1 binds tightly to heat shock proteins Hsp70/Hsc70, it was recognized that this new protein and other homologous members are regulators that modulate responses to cell death or cell stress through protein-protein interactions. BAG-1 and other members of the BAG family bind to Hsc70 through the conserved BAG domain. This domain binds to the ATPase domain of Hsc70. Recognizing the need for structural data to clearly define the binding interface beteen BAG-1 and Hsc70, we plan experiments to focus on this important protein-protein interaction. We have already made considerable progress on the NMR structure of BAG-1 in collaboration with David Hoyt at EMSL.Virtually complete HN, N Calpha, Cbeta,C' and Halpha assignments have been established. Now the next step is to gain structural insight into the molecular interactions at the binding interface between these two molecules. We plan to compare the chemical shifts in the free and bound form of BAG-1 when in complex with the Hsc70 protein. These experiments will localize the interface and map the residues on BAG-1 that are critical for binding to the molecular chaperone. Because the size of the complex of BAG-1 with the ATPase domain of Hsc70 approximates 60 kDa, TROSY and CRINEPT based experiments at high fields will be necessary. We also plan to employ the cross-saturation method which is documented to be useful for large complexes. These technically challenging experiments will require high field instruments. Therefore, we are requesting time on the 800 MHz spectrometer.
Project Details
Project type
Capability Research
Start Date
2000-09-29
End Date
2001-03-02
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members