Purification, crystallization and preliminary X-ray analysis of isocitrate dehydrogenase kinase/phosphatase from Escherichia coli.
The Escherichia coli aceK gene encodes isocitrate dehydrogenase kinase/phosphatase (EC 2.7.11.5), a bifunctional protein that phosphorylates and dephosphorylates isocitrate dehydrogenase (IDH), resulting in its inactivation and activation, respectively. This reversible (de)phosphorylation directs isocitrate, an intermediate of the citric acid cycle, to either go through the full cycle or to enter the glyoxylate bypass. In the present study, the AceK protein from E. coli has been purified and crystallized. Three crystal forms were obtained from very similar crystallization conditions. The crystals belong to space groups P4(1)2(1)2, P3(2)21 and P2(1)2(1)2(1) and diffracted X-rays to resolutions of 2.9, 3.0 and 2.7 A, respectively.
Zheng
Jimin
J
Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
Lee
Daniel C
DC
Jia
Zongchao
Z
D003460:Crystallization; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D007521:Isocitrate Dehydrogenase; D009097:Multienzyme Complexes / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism
D029968:Escherichia coli Proteins; D009097:Multienzyme Complexes; C548023:AceK protein, E coli; D007521:Isocitrate Dehydrogenase; D017346:Protein Serine-Threonine Kinases; C042301:isocitrate dehydrogenase kinase