杜氏利什曼原虫
生物化学
生物信息学
磷酸戊糖途径
生物
酶
活动站点
利什曼原虫
对接(动物)
水解酶
内脏利什曼病
基因
遗传学
利什曼病
护理部
万维网
寄生虫寄主
医学
计算机科学
糖酵解
作者
Tushar Lalaso Dhumal,Rajender Kumar,Anindita Paul,Pradyot Kumar Roy,Uttam Chand Banerjee,Sushma Singh
出处
期刊:Biochimie
[Elsevier]
日期:2022-08-01
卷期号:202: 212-225
标识
DOI:10.1016/j.biochi.2022.08.006
摘要
The enzymes of the pentose phosphate pathway are vital to survival in kinetoplastids. The second step of the pentose phosphate pathway involves hydrolytic cleavage of 6-phosphogluconolactone to 6-phosphogluconic acid by 6- phosphogluconolactonase (6PGL). In the present study, Leishmania donovani 6PGL (Ld6PGL) was cloned and overexpressed in bacterial expression system. Comparative sequence analysis revealed the conserved sequence motifs, functionally and structurally important residues in 6PGL family. In silico amino acid substitution study and interacting partners of 6PGL were predicted. The Ld6PGL enzyme was found to be active in the assay and in the parasites. Specificity was confirmed by Western blot analysis. The ∼30 kDa protein was found to be a dimer in MALDI, glutaraldehyde crosslinking and size exclusion chromatography studies. Kinetic analysis and structural stability studies of Ld6PGL were performed with denaturants and at varied temperature. Computational 3D Structural modelling of Ld6PGL elucidates that it has a similar α/β hydrolase fold structural topology as in other members of 6PGL family. The three loops are found in extended form when the structure is compared with the human 6PGL (Hs6PGL). Further, enzyme substrate binding mode and its mechanism were investigated using the molecular docking and molecular simulation studies. Interesting dynamics action of substrate 6-phosphogluconolactone was observed into active site during MD simulation. Interesting differences were observed between host and parasite enzyme which pointed towards its potential to be explored as an antileishmanial drug target. This study forms the basis for further analysis of the role of Ld6PGL in combating oxidative stress in Leishmania.
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