title: The PE16 (Rv1430) of Mycobacterium tuberculosis is an esterase belonging to serine hydrolase superfamily of proteins.
abstract: The PE and PPE multigene families, first discovered during the sequencing of M. tuberculosis H37Rv genome are responsible for antigenic variation and have been shown to induce increased humoral and cell mediated immune response in the host. Using the bioinformatics tools, we had earlier reported that the 225 amino acid residue PE-PPE domain (Pfam: PF08237) common to some PE and PPE proteins has a "serine α/β hydrolase" fold and conserved Ser, Asp and His catalytic triad characteristic of lipase, esterase and cutinase activities. In order to prove experimentally that PE-PPE domain is indeed a serine hydrolase, we have cloned the full-length Rv1430 and its PE-PPE domain into pET-28a vector, expressed the proteins in E. coli and purified to homogeneity. The activity assays of both purified proteins were carried out using p-nitrophenyl esters of aliphatic carboxylic acids with varying chain length (C2-C16) to study the substrate specificity. To characterize the active site of the PE-PPE domain, we mutated the Ser199 to Ala. The activity of the protein in the presence of serine protease inhibitor- PMSF and the mutant protein were measured. Our results reveal that Rv1430 and its PE-PPE domain possess esterase activity and hydrolyse short to medium chain fatty acid esters with the highest specific activity for pNPC6 at 37°C, 38°C and pH 7.0, 8.0. The details of this work and the observed results are reported in this manuscript.
authors: School of Chemistry, University of Hyderabad, Hyderabad, India.
authors: Rafiya
authors: R
authors: Sultana
authors: Mani Harika
authors: MH
authors: Vemula
authors: Sharmishta
authors: S
authors: Banerjee
authors: Lalitha
authors: L
authors: Guruprasad
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D015153:Blotting, Western; D020134:Catalytic Domain / Q000235:genetics; D002942:Circular Dichroism; D003001:Cloning, Molecular; D019295:Computational Biology; D017931:DNA Primers / Q000235:genetics; D004926:Escherichia coli; D004950:Esterases / Q000235:genetics*; D006863:Hydrogen-Ion Concentration; D005810:Multigene Family / Q000235:genetics*; D009169:Mycobacterium tuberculosis / Q000201:enzymology* / Q000235:genetics; D012697:Serine Endopeptidases / Q000235:genetics*; D013696:Temperature
chemical_list: D001426:Bacterial Proteins; D017931:DNA Primers; D004950:Esterases; D012697:Serine Endopeptidases
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