Crystallization and characterization of polyphosphate kinase from Escherichia coli.
Linear polyphosphate chains have been found to play a key role in bacterial responses to stresses and nutritional depletion, and are necessary for host infection of various pathogens. Polyphosphate kinase (PPK) is a critical enzyme responsible for polyphosphate synthesis in bacteria. PPK knockout mutations in several Gram-negative pathogens identify PPK as an ideal drug target for the development of a new class of antibacterial drugs. To reveal the catalytic mechanism and provide a structural basis for drug discovery, we have purified and crystallized full-length Escherichia coli PPK and its complex with AMP-PNP. The crystals diffract to a resolution of 2.5A and belong to the space group P4(2)2(1)2 with unit-cell parameters a=152.0, b=152.0, and c=150.0 A. Crystal structure of PPK is being determined by the Se-Met MAD experiment.
Zhu
Ye
Y
Department of Biological Structure, University of Washington, Seattle, WA 98195, USA.
Lee
Sam S K
SS
Xu
Wenqing
W
D000266:Adenylyl Imidodiphosphate / Q000378:metabolism; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology*; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D017853:Phosphotransferases (Alcohol Group Acceptor) / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D017856:Phosphotransferases (Phosphate Group Acceptor) / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D012995:Solubility
D029968:Escherichia coli Proteins; D000266:Adenylyl Imidodiphosphate; D017853:Phosphotransferases (Alcohol Group Acceptor); D017856:Phosphotransferases (Phosphate Group Acceptor); C022424:polyphosphate kinase; C475691:polyphosphate kinase, E coli