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Updated: Jun 23, 2026

A Purification and In Vitro Activity Assay for a (p)ppGpp Synthetase from Clostridium difficile
Published on: November 3, 2018
Purification, crystallization and preliminary X-ray analysis of isocitrate dehydrogenase kinase/phosphatase from
Jimin Zheng1, Daniel C Lee, Zongchao Jia
1Department of Biochemistry, Queen's University, Kingston, Ontario, Canada.
Abstract:
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.

