Metal-based Enzyme Promiscuity Of Mammalian Senescence Marker Protein-30 | 38963
Journal of Biotechnology & Biomaterials
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Senescence marker protein-30 (SMP30) is a lactonase enzyme involved in the biosynthesis of ascorbic acid in mice. However
in human, there is no ascorbic acid biosynthesis, the expression of SMP30 has been reported in liver, kidney and other
tissues. Hence, to study the role of SMP30 in human is an area of intensive research. SMP30 is also known as Regucalcin due
to its involvement in calcium homeostasis. Recent studies showed its association in organophosphate hydrolysis including
nerve agents like sarin and soman. Since SMP30 is also involved in calcium homeostasis, we are investigating the metal-based
substrate specificities shown by SMP30. Studying the enzymatic activities and metal-based substrate specificities of SMP30 in
the presence of different divalent cations may be a useful approach for discovering the precise physiological role in human.
Therefore, cDNA of mouse liver SMP30 was cloned in bacterial expression vector and sequence verified. The protein was
overexpressed with 6xHis tag in E. coli (BL21) cells and purified for biochemical characterization. We used a bacterial codonoptimized
synthetic gene to study the human SMP30. Purified proteins were used to measure the activities in the presence
of different metals like Ca2+, Cd2+, Co2+, Mg2+ and Mn2+. Metal specificities were examined with several substrates like phenyl
acetate, naphthyl acetate, parathiol, paraoxon and thiobutyrolacton. SMP30 shows highest esterase activity in the presence of
Co2+ as compare to other divalent cations.
Rinkoo Devi Gupta has completed her PhD from Banaras Hindu University and Postdoctoral studies from Weizmann Institute of Sciences, Israel. She is working as an Assistant Professor in South Asian University, New Delhi, an international university established by SAARC nations. She has published more than 25 papers in reputed journals including Nature Chemical Biology and Nature Methods. Her area of current research is to understand the molecular evolution of enzymes, protein engineering and drug discoveries.