对耐药和敏感临床分离株中吡嗪酰胺酶和DNA旋转酶蛋白质结构的见解。（原文中“of.”后面似乎缺少具体内容）

Insights into Pyrazinamidase and DNA Gyrase Protein Structures in Resistant and Susceptible Clinical Isolates of .

作者信息

Ahmadi Azam, Nazari Raziyeh, Arjomandzadegan Mohammad, Zolfaghari Mohammad Reza, Vahidi Vahideh, Poolad Toktam, Kahbazi Manijeh, Sadrnia Maryam, Tousheh Mojtaba, Rafiee Pourya

机构信息

Department of Molecular Genetics, Tarbiat Modares University,; Infectious Diseases Research Center (IDRC), Arak University of Medical Sciences, Arak, Iran.

Department of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.

出版信息

Tanaffos. 2016;15(3):147-153.

PMID:28210279

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC5304958/

Abstract

BACKGROUND

Mutations in and genes cause pyrazinamide (PZA) and fluroquinolone resistance in (). In the present study, structures of pyrazinamidase (PZase) and DNA gyrase proteins were studied in resistant and susceptible clinical isolates of

MATERIALS AND METHODS

Sixty clinical isolates of were used in this study. Polymerase chain reaction (PCR) amplification of and genes was accomplished on purified DNA. Sequence of the fragments was determined by an Applied BiosystemsTM apparatus. Bioinformatic analysis was performed by online software and three-dimensional (3D) structures of proteins was predicted using Molegro Virtual Docker (MVD) Modeler software.

RESULTS

Amplified 744 and 194 bp fragments of and genes, respectively were yielded suitable sequence results. Predicted 3D structures of proteins showed some differences between wild-type and mutant structures. Mutation in amino acid No.31 (T92C) caused an increase in distance from metal ion position to enzyme active site, but it was considered as a polymorphism. Docking results by MVD revealed a relationship in quinolone resistance-determining regions (QRDR) amino acids in interaction with antibiotic. T92C mutation in PZase from non-polar aliphatic amino acid Ile (ATC) to polar aliphatic amino acid threonine (ACC) was a polymorphism.

CONCLUSION

Structural changes in two important proteins related to drug resistance were proven in clinical isolates of .

摘要

背景

结核分枝杆菌中的pncA和gyrA基因发生突变会导致对吡嗪酰胺（PZA）和氟喹诺酮耐药。在本研究中，对结核分枝杆菌耐药和敏感临床分离株中的吡嗪酰胺酶（PZase）和DNA促旋酶蛋白结构进行了研究。

材料与方法

本研究使用了60株结核分枝杆菌临床分离株。在纯化的DNA上完成pncA和gyrA基因的聚合酶链反应（PCR）扩增。片段序列由Applied BiosystemsTM仪器测定。通过在线软件进行生物信息学分析，并使用Molegro Virtual Docker（MVD）建模软件预测蛋白质的三维（3D）结构。

结果

分别扩增出744 bp和194 bp的pncA和gyrA基因片段，获得了合适的序列结果。预测的蛋白质3D结构显示野生型和突变型结构之间存在一些差异。31号氨基酸（T92C）的突变导致金属离子位置到酶活性位点的距离增加，但被认为是一种多态性。MVD对接结果显示喹诺酮耐药决定区（QRDR）氨基酸与抗生素相互作用之间存在关系。PZase中T92C突变使非极性脂肪族氨基酸异亮氨酸（ATC）变为极性脂肪族氨基酸苏氨酸（ACC），这是一种多态性。