Please use this identifier to cite or link to this item: http://nopr.niscair.res.in/handle/123456789/56611
Title: Cloning, expression and purification of alkylhydroperoxidase C, a crucial respiratory antioxidant in Brucella abortus
Authors: Goel, Divya
Kumar, Nitesh
Ghosh, Debasish
Kumar, Sudhir
Keywords: Alkylhydroperoxidase C;Brucella abortus;BaAhpC gene cloning
Issue Date: Oct-2020
Publisher: NISCAIR-CSIR, India
Abstract: 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.
Page(s): 273-281
URI: http://nopr.niscair.res.in/handle/123456789/56611
ISSN: 0975-0967 (Online); 0972-5849 (Print)
Appears in Collections:IJBT Vol.19(4) [October 2020]

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