PMID: 9647837Jul 2, 1998Paper

Cloning and expression of the inositol monophosphatase gene from Methanococcus jannaschii and characterization of the enzyme

Applied and Environmental Microbiology
L Chen, Mary F Roberts

Abstract

Inositol monophosphatase (EC 3.1.3.25) plays a pivotal role in the biosynthesis of di-myo-inositol-1,1'-phosphate, an osmolyte found in hyperthermophilic archaeal. Given the sequence homology between the MJ109 gene product of Methanococcus jannaschii and human inositol monophosphatase, the MJ109 gene was cloned and expressed in Escherichia coli and examined for inositol monophosphatase activity. The purified MJ109 gene product showed inositol monophosphatase activity with kinetic parameters (K(m) = 0.091 +/- 0.016 mM; Vmax = 9.3 +/- 0.45 mumol of Pi min-1 mg of protein-1) comparable to those of mammalian and E. coli enzymes. Its substrate specificity, Mg2+ requirement, Li+ inhibition, subunit association (dimerization), and heat stability were studied and compared to those of other inositol monophosphatases. The lack of inhibition by low concentrations of Li+ and high concentrations of Mg2+ and the high rates of hydrolysis of glucose-1-phosphate and p-nitrophenylphosphate are the most pronounced differences between the archaeal inositol monophosphatase and those from other sources. The possible causes of these kinetic differences are discussed, based on the active site sequence alignment between M. jannaschii and human inositol...Continue Reading

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Citations

Apr 10, 2002·The Journal of Biological Chemistry·Kimberly A StieglitzBoguslaw Stec
Sep 30, 2005·The Journal of Biological Chemistry·Shigeyuki KawaiKousaku Murata
Mar 16, 2007·Proceedings of the National Academy of Sciences of the United States of America·Dmitry A RodionovAndrei L Osterman
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Feb 20, 2016·Acta Crystallographica. Section D, Structural Biology·Sudipta BhattacharyyaAmit Kumar Das
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Jan 16, 2009·Journal of Medicinal Chemistry·Aleksandra RudnitskayaKimberly Stieglitz
Dec 6, 2005·Biochemical and Biophysical Research Communications·Xiaoling GuHonghai Wang

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