7E4L

Conversion of pyrophosphate-dependent myo-inositol-1 kinase into myo-inositol-3 kinase by N78L/S89L mutation

Summary for 7E4L

• Entry DOI: 10.2210/pdb7e4l/pdb
• Descriptor: PfkB domain-containing protein, METHYLENEDIPHOSPHONIC ACID, MAGNESIUM ION, ... (4 entities in total)
• Functional Keywords: pyrophosphate-dependent kinase, transferase
• Biological source: Thermotoga maritima (strain ATCC 43589 / MSB8 / DSM 3109 / JCM 10099)
• Total number of polymer chains: 2
• Total formula weight: 64660.19

Authors

Tashiro, R.,Miki, K.,Fujihashi, M. (deposition date: 2021-02-14, release date: 2021-05-05, Last modification date: 2023-11-29)

Primary citation

Tashiro, R.,Sato, T.,Atomi, H.,Miki, K.,Fujihashi, M.
Altering the Phosphorylation Position of Pyrophosphate-Dependent myo -Inositol-1-Kinase Based on Its Crystal Structure.
Acs Chem.Biol., 16:794-799, 2021

PubMed Abstract: 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.

PubMed: 33877806
DOI: 10.1021/acschembio.0c00733

Experimental method

X-RAY DIFFRACTION (1.6 Å)