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{sub 1}2{sub 1}2{sub 1}, with unit-cell parameters a = 117.1, b = 127.7, c = 170.7 {angstrom}. A complete X-ray intensity data set has been collected and processed to 2.50 {angstrom} 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.
- Research Organization:
- SLAC National Accelerator Lab., Menlo Park, CA (United States). Stanford Synchrotron Radiation Lightsource (SSRL)
- Sponsoring Organization:
- USDOE Office of Science (US)
- DOE Contract Number:
- AC03-76SF00515
- OSTI ID:
- 815900
- Report Number(s):
- SLAC-REPRINT-2002-305; TRN: US200319%%574
- Journal Information:
- Acta Crystallographica Section D, Other Information: Acta Crystallogr.D Biol.Crystallogr. 58:515,2002; PBD: 1 Jan 2002
- Country of Publication:
- United States
- Language:
- English
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