Large-scale crystallization and neutron crystallographic analysis of HSP70 in complex with ADP.
HSP70 belongs to the heat-shock protein family and binds to unfolded proteins, driven by ATP hydrolysis, in order to prevent aggregation. Previous X-ray crystallographic analyses of HSP70 have shown that HSP70 binds to ADP with internal water molecules. In order to elucidate the role of the water molecules, including their H/D atoms, a neutron diffraction study of the human HSP70 ATPase domain was initiated. Deuterated large crystals of the HSP-ADP complex (1.2-1.8 mm3) were successfully grown by large-scale crystallization, and a neutron diffraction experiment at BIODIFF resulted in diffraction to a maximum resolution of 2.2 Å. After data reduction, the overall completeness, Rmeas and average I/σ(I) were 90.4%, 11.7% and 8.1, respectively, indicating that the data set was sufficient to visualize H and D atoms.
['Yokoyama|Takeshi|T|', 'Ostermann|Andreas|A|', 'Schrader|Tobias E|TE|', 'Mizuguchi|Mineyuki|M|']
D000244:Adenosine Diphosphate D000595:Amino Acid Sequence D003460:Crystallization D018360:Crystallography, X-Ray D018840:HSP70 Heat-Shock Proteins D006801:Humans D033363:Neutron Diffraction D014961:X-Ray Diffraction
large-scale crystallization HSP70 heat-shock proteins neutron protein crystallography