Altering the phosphorylation position of pyrophosphate-dependent myo-inositol-1-kinase based on its crystal structure
Altering the phosphorylation position of pyrophosphate-dependent myo-inositol-1-kinase based on its crystal structure
复制标题
根据焦磷酸依赖性肌醇-1-激酶的晶体结构改变其磷酸化位置
DOI：
10.1021/acschembio.0c00733
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发表时间：
2021
期刊：
影响因子：
4
通讯作者：
M.
中科院分区：
文献类型：
--
作者：
Tashiro;R.;Sato;T.;Atomi;H.;Miki;K.;Fujihashi;M.
大多数激酶利用ATP作为磷酸供体，并使多种磷酸受体磷酸化。另一种磷酸盐供体是无机焦磷酸盐（PPi），其成本仅为ATP的1/1000。为了开发一种设计ppi依赖性激酶的方法，我们在此旨在将ppi依赖性肌醇激酶的产物从-肌醇- 1-磷酸改为-肌醇- 3-磷酸。为此，我们将atp依赖性肌肌醇-3激酶的肌肌醇识别残基引入到ppi依赖性肌肌醇-1激酶中。这种取代有望改变肌醇在活性位点的三维排列，并使3C位置的羟基靠近催化残基。LC-MS和NMR分析证实，该工程酶成功地从肌醇和肌醇中分离出肌醇3-磷酸。
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-dependentmyo-inositol kinase fromd-myo-inositol 1-phosphate tod-myo-inositol 3-phosphate. For this purpose, we introduced themyo-inositol recognition residues of the ATP-dependentmyo-inositol-3-kinase into the PPi-dependentmyo-inositol-1-kinase. This replacement was expected to change the 3D arrangements ofmyo-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 producedmyo-inositol 3-phosphate from PPi andmyo-inositol.