Cloning and characterization of GTP-binding proteins of Mycobacterium tuberculosis H(37)Rv

Enzyme and Microbial Technology
Laxman S MeenaY Singh

Abstract

GTP-binding proteins (G-proteins) are highly conserved signaling molecules that participate in cellular signaling and bacterial pathogenesis by regulating the activity of cognate GTPases. However, the exact role of G-proteins in the pathogenesis of Mycobacterium tuberculosis is poorly understood. The complete genome sequence of M. tuberculosis H(37)Rv, suggests the presence of several homologs of bacterial G-proteins. In the present study, three G-proteins, Era, Obg and LepA of M. tuberculosis H(37)Rv were cloned and expressed in Escherichia coli. Purified proteins showed GTP-binding and hydrolyzing activities. A point mutation in the conserved GTP-binding motif, AspXXGly (Asp to Ala) in Era (Asp-258) and Obg (Asp-212) proteins resulted in the loss of the associated activities, confirming that known key residues in well-established G-proteins are also conserved in mycobacterial homologs. This study confirms that Era, Obg and LepA of M. tuberculosis H(37)Rv possess GTPase activity and provide a platform to understand the physiological significance of these proteins in associated pathogenesis.

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Citations

Mar 30, 2013·Critical Reviews in Microbiology·Cyrielle KintJan Michiels
Feb 19, 2011·Biologicals : Journal of the International Association of Biological Standardization·Laxman S Meena, Rajni
Jun 18, 2010·The FEBS Journal·Laxman S Meena, Rajni
Mar 9, 2010·International Journal of Infectious Diseases : IJID : Official Publication of the International Society for Infectious Diseases· Rajni, Laxman S Meena
Jan 8, 2021·International Journal of Biological Macromolecules· ShivangiLaxman S Meena

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