6K5L

The crystal structure of isocitrate dehydrogenase kinase/phosphatase wtih two Mn2+ from E. coli

Summary for 6K5L

• Entry DOI: 10.2210/pdb6k5l/pdb
• Descriptor: Isocitrate dehydrogenase kinase/phosphatase, ADENOSINE MONOPHOSPHATE, ADENOSINE-5'-DIPHOSPHATE, ... (5 entities in total)
• Functional Keywords: kinase, phosphatase, hydrolase
• Biological source: Escherichia coli O157
• Total number of polymer chains: 2
• Total formula weight: 137415.71

Authors

Zhang, X.,Lei, Z.,Zheng, J.,Jia, Z. (deposition date: 2019-05-29, release date: 2019-07-03, Last modification date: 2023-11-22)

Primary citation

Zhang, X.,Shen, Q.,Lei, Z.,Wang, Q.,Zheng, J.,Jia, Z.
Characterization of metal binding of bifunctional kinase/phosphatase AceK and implication in activity modulation.
Sci Rep, 9:9198-9198, 2019

PubMed Abstract: A unique bifunctional enzyme, isocitrate dehydrogenase kinase/phosphatase (AceK) regulates isocitrate dehydrogenase (IDH) by phosphorylation and dephosphorylation in response to nutrient availability. Herein we report the crystal structure of AceK in complex with ADP and Mn ions. Although the overall structure is similar to the previously reported structures which contain only one Mg ion, surprisingly, two Mn ions are found in the catalytic center of the AceK-Mn structure. Our enzymatic assays demonstrate that AceK-Mn showed higher phosphatase activity than AceK-Mg, whereas the kinase activity was relatively unaffected. We created mutants of AceK for all metal-coordinating residues. The phosphatase activities of these mutants were significantly impaired, suggesting the pivotal role of the binuclear (M1-M2) core in AceK phosphatase catalysis. Moreover, we have studied the interactions of Mn and Mg with wild-type and mutant AceK and found that the number of metal ions bound to AceK is in full agreement with the crystal structures. Combined with the enzymatic results, we demonstrate that AceK exhibits phosphatase activity in the presence of two, but not one, Mn ions, similar to PPM phosphatases. Taken together, we suggest that metal ions help AceK to balance and fine tune its kinase and phosphatase activities.

PubMed: 31235769
DOI: 10.1038/s41598-019-45704-z

Experimental method

X-RAY DIFFRACTION (2.55 Å)