title: Crystallization, preliminary X-ray analysis and biophysical characterization of HPr kinase/phosphatase of Mycoplasma pneumoniae.
abstract: 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.
authors.affiliation: Lehrstuhl für Mikrobiologie, Friedrich-Alexander-Universität Erlangen-Nürnberg, Staudtstrasse 5, D-91058 Erlangen, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Steinhauer
authors.forename: Gregory S
authors.initials: GS
authors.lastname: Allen
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Hillen
authors.forename: Jörg
authors.initials: J
authors.lastname: Stülke
authors.forename: Richard G
authors.initials: RG
authors.lastname: Brennan
mesh_terms: D001426:Bacterial Proteins; D003460:Crystallization; D018360:Crystallography, X-Ray; D004228:Dithionitrobenzoic Acid / Q000737:chemistry; D008958:Models, Molecular; D009177:Mycoplasma pneumoniae / Q000737:chemistry / Q000201:enzymology*; D011487:Protein Conformation; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry*; D017186:Titrimetry
chemical_list: D001426:Bacterial Proteins; D004228:Dithionitrobenzoic Acid; C110847:HPr kinase; D017346:Protein Serine-Threonine Kinases