Expression and purification of a protein kinase, PknH of Mycobacterium bovis BCG pasteur 1173P2

Lau, Sing Leng (2005) Expression and purification of a protein kinase, PknH of Mycobacterium bovis BCG pasteur 1173P2. Universiti Malaysia Sabah. (Unpublished)

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Abstract

Tuberculosis (TB) is a mycobacterial disease caused by the pathogenic Mycobacterium tuberculosis. The survival of the tubercle bacilli in the intracellular environment of macrophage causes prolonged latent infection of the disease. Pathogenesis of M. tuberculosis is closely connected to its extracellular stresses adaptation, as well as intracellular survival within the host. Signal transduction through protein phosphorylation has long been emphasized in their relationship with bacterial pathogenesis. These regulatory mechanism have shown to be mediated by the universal regulatory protein known as Serine/Threonine Protein Kinases (STPKs). By then, molecular approach has opened a new leaf in the battle against TB. The present study investigates one of the STPKs from an attenuated strain of Mycobacterium, which is PknH of Mycobacterium bovis BCG Pasteur 1173P2. The pknH gene from M. bovis BCG Pasteur 1173P2 genome was cloned and expressed under IPTG-induction as Histidine tagged recombinant protein in Escherichia coli. The recombinant protein was purified from the insoluble fraction by affinity chromatography using Ni²�-NTA resin. SDS-PAGE revealed the presence of a full-length Histidine-tagged PknH at a position of 66 kDa in size. The concentration of purified PknH was detennined. Analysis of the nucleotide-derived amino acid sequence demonstrated that PknH is a transmembrane eukaryotic-like (Hank's-type) STPK harbouring all 11 subdomains. Sequence alignment of PknH with other bacterial STPKs showing similarity between these bacteria, with the subdomains highly conserved. The closest homologues are none other than the member of the same genus, Mycobacterium. Possible roles of PknH in processes that are unique to the pathogenic strains and their environmental sensing mechanism have been proposed. Involvement of PknH in the signaling pathway has been discussed, particularly in the regulation of cell wall arabinan metabolism

Item Type: Academic Exercise
Uncontrolled Keywords: Tuberculosis, tubercle bacilli, Serine/Threonine Protein Kinases, PknH, Mycobacterium bovis BCG Pasteur 1173P2
Subjects: Q Science > QH Natural history > QH301 Biology
Divisions: SCHOOL > School of Science and Technology
Depositing User: MDM SITI AZIZAH IDRIS
Date Deposited: 27 Feb 2014 06:57
Last Modified: 12 Oct 2017 05:27
URI: http://eprints.ums.edu.my/id/eprint/8367

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