Role of a disulphide bond in Helicobacter pylori arginase |
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Authors: | Abhishek Srivastava |
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Institution: | National Institute of Immunology, Aruna Asaf Ali Marg, New Delhi 110 067, India |
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Abstract: | Arginase is a binuclear Mn2+-metalloenzyme of urea cycle that hydrolyses arginine to ornithine and urea. Unlike other arginases, the Helicobacter pylori enzyme is selective for Co2+. Previous study reported that DTT strongly inhibits the H. pylori enzyme activity suggesting that a disulphide bond is critical for the catalysis. In this study, we have undertaken steady-state kinetics, circular dichroism and mutational analysis to examine the role of a disulphide bond in this protein. By mutational analysis, we show that the disulphide bond is not important for catalytic activity; rather it plays an important role for the stability of the protein as observed from thermal denaturation studies. The loss of catalytic activity in the wild-type protein with DTT is due to the interaction with Co2+. This is verified with the Mn2+-reconstituted proteins which showed a marginal loss in the activity with DTT. |
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Keywords: | H pylori arginase Disulphide bond Thermal stability Co(II) CD |
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