Altering the Phosphorylation Position of Pyrophosphate-Dependent myo -Inositol-1-Kinase Based on Its Crystal Structure

Most kinases utilize ATP as a phosphate donor and phosphorylate a wide range of phosphate acceptors. An alternative phosphate donor is inorganic pyrophosphate (PPi), which costs only 1/1000 of ATP. To develop a method to engineer PPi-dependent kinases, we herein aimed to alter the product of PPi-dep...

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Published inACS chemical biology Vol. 16; no. 5; pp. 794 - 799
Main Authors Tashiro, Ryo, Sato, Takaaki, Atomi, Haruyuki, Miki, Kunio, Fujihashi, Masahiro
Format Journal Article
LanguageEnglish
Published United States 21.05.2021
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Summary:Most kinases utilize ATP as a phosphate donor and phosphorylate a wide range of phosphate acceptors. An alternative phosphate donor is inorganic pyrophosphate (PPi), which costs only 1/1000 of ATP. To develop a method to engineer PPi-dependent kinases, we herein aimed to alter the product of PPi-dependent -inositol kinase from d- inositol 1-phosphate to d- inositol 3-phosphate. For this purpose, we introduced the inositol recognition residues of the ATP-dependent inositol-3-kinase into the PPi-dependent -inositol-1-kinase. This replacement was expected to change the 3D arrangements of -inositol in the active site and bring the hydroxyl group at the 3C position close to the catalytic residue. LC-MS and NMR analyses proved that the engineered enzyme successfully produced inositol 3-phosphate from PPi and inositol.
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ISSN:1554-8929
1554-8937
DOI:10.1021/acschembio.0c00733