Assay development and inhibition of the Mt-DprE2 essential reductase from Mycobacterium tuberculosis.
Assay development and inhibition of the Mt-DprE2 essential reductase from Mycobacterium tuberculosis.
复制标题
DOI：
10.1099/mic.0.001288
复制
发表时间：
2023-01
期刊：
影响因子：
2.8
通讯作者：
Besra, Gurdyal S.
中科院分区：
文献类型：
--
作者：
Batt, Sarah M.;Toth, Szilvi;Rodriguez, Beatriz;Abrahams, Katherine A.;Veerapen, Natacha;Chiodarelli, Giacomo;Cox, Liam R.;Moynihan, Patrick J.;Lelievre, Joel;Futterer, Klaus;Besra, Gurdyal S.
关键词：
DprE 2是合成十异戊二烯基磷酰基-β-d-阿拉伯呋喃糖（DPA）和随后的阿拉伯半乳聚糖的必需酶，并且是治疗糖尿病的重要新药靶点。 M.结核通过Mt-DprE 2-靶标过表达研究鉴定了GSK-177盒组中的两种化合物GSK 301 A和GSK 032 A。共纯化Mt-DprE 1-DprE 2复合物，并基于DprE 2的还原型烟酰胺腺嘌呤二核苷酸辅因子（NADH/NADPH）的氧化，开发了一种新的体外DprE 2测定法。Mt-DprE 1-DprE 2复合物在基于DprE 1刃天青的试验中显示出有趣的动力学，其中发现Mt-DprE 2增强Mt-DprE 1活性并降低底物抑制;在DprE 2试验中也显示出类似的底物抑制和两种潜在辅因子NADH和NADPH的动力学差异。尽管在DprE 2试验中未观察到两种GSK组化合物的抑制作用，但自发突变体的产生表明可能存在前药活化途径形式的解释，涉及fgd 1和fbiC。
DprE2 is an essential enzyme in the synthesis of decaprenylphosphoryl-β-d-arabinofuranose (DPA) and subsequently arabinogalactan, and is a significant new drug target for M. tuberculosis . Two compounds from the GSK-177 box set, GSK301A and GSK032A, were identified through Mt-DprE2-target overexpression studies. The Mt-DprE1-DprE2 complex was co-purified and a new in vitro DprE2 assay developed, based on the oxidation of the reduced nicotinamide adenine dinucleotide cofactor of DprE2 (NADH/NADPH). The Mt-DprE1-DprE2 complex showed interesting kinetics in both the DprE1 resazurin-based assay, where Mt-DprE2 was found to enhance Mt-DprE1 activity and reduce substrate inhibition; and also in the DprE2 assay, which similarly exhibited substrate inhibition and a difference in kinetics of the two potential cofactors, NADH and NADPH. Although, no inhibition was observed in the DprE2 assay by the two GSK set compounds, spontaneous mutant generation indicated a possible explanation in the form of a pro-drug activation pathway, involving fgd1 and fbiC.
登录
查看更多内容
DOI：
10.1073/pnas.0508392103
发表时间：
2006-01-10
影响因子：
11.1
作者：
Manjunatha, UH;Boshoff, H;Barry, CE
通讯作者：
Barry, CE
影响因子：
3.7
作者：
Jose Rebollo-Lopez, Maria;Lelievre, Joel;Barros-Aguire, David
通讯作者：
Barros-Aguire, David
影响因子：
3.6
作者：
Kolly, Gaelle S.;Boldrin, Francesca;Cole, Stewart T.
通讯作者：
Cole, Stewart T.
影响因子：
4.4
作者：
Carroll, Paul;Faray-Kele, Marie-Claire;Parish, Tanya
通讯作者：
Parish, Tanya
影响因子：
3.7
作者：
Crellin, Paul K.;Brammananth, Rajini;Coppel, Ross L.
通讯作者：
Coppel, Ross L.