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Altering the Phosphorylation Position of Pyrophosphate-Dependent <i>myo</i>-Inositol-1-Kinase Based on Its Crystal Structure
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- Ryo Tashiro
- Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
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- Takaaki Sato
- Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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- Haruyuki Atomi
- Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto 615-8510, Japan
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- Kunio Miki
- Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
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- Masahiro Fujihashi
- Department of Chemistry, Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto 606-8502, Japan
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Description
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 myo-inositol kinase from d-myo-inositol 1-phosphate to d-myo-inositol 3-phosphate. For this purpose, we introduced the myo-inositol recognition residues of the ATP-dependent myo-inositol-3-kinase into the PPi-dependent myo-inositol-1-kinase. This replacement was expected to change the 3D arrangements of myo-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 myo-inositol 3-phosphate from PPi and myo-inositol.
Journal
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- ACS Chemical Biology
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ACS Chemical Biology 16 (5), 794-799, 2021-04-20
American Chemical Society (ACS)
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Keywords
- 571
- Physiology
- Plant Biology
- 3 D arrangements
- Biochemistry
- Microbiology
- 3 C position
- alternative phosphate donor
- Molecular Biology
- engineer PPi-dependent kinases
- Pharmacology
- inositol 1- phosphate
- PPi-dependent myo
- Cell Biology
- NMR
- LC-MS
- ATP
- Mental Health
- inositol 3- phosphate
- inositol recognition residues
- Neuroscience
- Biotechnology
- Developmental Biology
Details 詳細情報について
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- CRID
- 1360009142872660992
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- ISSN
- 15548937
- 15548929
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- Article Type
- journal article
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- Data Source
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- Crossref
- KAKEN
- OpenAIRE
