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Cloning, expression and purification of alkylhydroperoxidase C, a crucial respiratory antioxidant in Brucella abortus

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Title Cloning, expression and purification of alkylhydroperoxidase C, a crucial respiratory antioxidant in Brucella abortus
 
Creator Goel, Divya
Kumar, Nitesh
Ghosh, Debasish
Kumar, Sudhir
 
Subject Alkylhydroperoxidase C
Brucella abortus
BaAhpC gene cloning
 
Description 273-281
During infection, Brucellae face extensive oxidative pressure inside host macrophages in the form of oxidative burst and reactive oxygen species (ROS). Alkylhydroperoxidase C (AhpC) is the primary scavenger of these ROS in Brucella and alkylhydroperoxidase D (AhpD) helps it to regain its reduced state after the catalytic reaction. In this study, we attempted to crystallize the AhpC protein from B. abortus. Extensive crystallization attempts were carried out but crystals could not be obtained. B. abortus AhpC (BaAhpC) gene was cloned and expressed in Escherichia coli, followed by protein purification using affinity and gel filtration chromatography. BaAhpC was found to be a dimer in solution. Protein sequence alignment analysis shows that BaAhpC share about 50% identity with Mycobacterium tuberculosis AhpC (MtbAhpC) protein and has 3 conserved cysteine residues which are directly involved in catalytic activity. The homology model of BaAhpC structure was prepared using MtbAhpC as a template which show it may exist in the form a dodecamer.
 
Date 2021-04-07T06:59:13Z
2021-04-07T06:59:13Z
2020-10
 
Type Article
 
Identifier 0975-0967 (Online); 0972-5849 (Print)
http://nopr.niscair.res.in/handle/123456789/56611
 
Language en_US
 
Rights CC Attribution-Noncommercial-No Derivative Works 2.5 India
 
Publisher NISCAIR-CSIR, India
 
Source IJBT Vol.19(4) [October 2020]