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.

Source: Acta Crystallographica Section F - September 22, 2017 Category: Biochemistry Authors: Yokoyama, T. Ostermann, A. Schrader, T.E. Mizuguchi, M. Tags: neutron protein crystallography heat-shock proteins HSP70 large-scale crystallization research communications Source Type: research