Crystallization, preliminary X-ray analysis and biophysical characterization of HPr kinase/phosphatase of Mycoplasma pneumoniae.
The Mycoplasma pneumoniae HPr kinase/phosphatase (HPrK/P) is a member of a large family of enzymes which are central to carbon regulation in Gram-positive bacteria. The full-length M. pneumonia HPrK/P was crystallized from solutions of polyethylene glycol 8000 and KCl or NaCl which also contained the non-hydrolysable ATP analog adenosine 5'-[beta, gamma-methylene]triphosphate (AMPPCP). The crystals belong to the orthorhombic space group P2(1)2(1)2(1), with unit-cell parameters a = 117.1, b = 127.7, c = 170.7 A. A complete X-ray intensity data set has been collected and processed to 2.50 A resolution. The slow self-rotation function revealed the presence of a sixfold axis. Dynamic light-scattering (DLS) experiments indicated a molecular weight of 197 kDa for HPrK/P in the absence of AMPPCP and of 217 kDa in the presence of the ATP analog. Thus, the biophysical and crystallographic data suggest that HPrK/P is a functional hexamer that undergoes an ATP-binding-induced conformational change.
Steinhauer, K; Allen, GS; Hillen, W; Stülke, J; Brennan, RG
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