Altering the Phosphorylation Position of Pyrophosphate-Dependent myo-Inositol-1-Kinase Based on Its Crystal Structure
| Autor: | Ryo Tashiro, Haruyuki Atomi, Masahiro Fujihashi, Takaaki Sato, Kunio Miki |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
0301 basic medicine
chemistry.chemical_classification biology 010405 organic chemistry Kinase Active site General Medicine Phosphate 01 natural sciences Biochemistry Pyrophosphate 0104 chemical sciences carbohydrates (lipids) 03 medical and health sciences Residue (chemistry) chemistry.chemical_compound 030104 developmental biology Enzyme chemistry biology.protein Molecular Medicine Phosphorylation lipids (amino acids peptides and proteins) Inositol |
| Zdroj: | ACS Chemical Biology. 16:794-799 |
| ISSN: | 1554-8937 1554-8929 |
| DOI: | 10.1021/acschembio.0c00733 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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