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KMID : 0624620110440120805
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BMB Reports
2011 Volume.44 No. 12 p.805 ~ p.810
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Characterization of a novel methionine sulfoxide reductase A from tomato (Solanum lycopersicum), and its protecting role in Escherichia coli
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Dai Changbo
Singh Naresh Kumar
Park Myung-Ho
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Abstract
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Methionine sulfoxide reductase A (MSRA) is a ubiquitous enzyme that has been demonstrated to reduce the S enantiomer of methionine sulfoxide (MetSO) to methionine (Met) and can protect cells against oxidative damage. In this study, we isolated a novel MSRA (SlMSRA2) from Micro-Tom (Solanum lycopersicum L. cv. Micro-Tom) and characterized it by subcloning the coding sequence into a pET expression system. Purified recombinant protein was assayed by HPLC after expression and refolding. This analysis revealed the absolute specificity for methionine-S-sulfoxide and the enzyme was able to convert both free and protein-bound MetSO to Met in the presence of DTT. In addition, the optimal pH, appropriate temperature, and Km and Kcat values for MSRA2 were observed as 8.5, 25oC, 352 ± 25 μM, and 0.066 ± 0.009 S(-1), respectively. Disk inhibition and growth rate assays indicated that SlMSRA2 may play an essential function in protecting E. coli against oxidative damage.
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KEYWORD
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Kinetics parameters, Micro-Tom, MSRA, Oxidative damage, Reverse phase HPLC
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