Isolation, purification and kinetic characterization of plasma membrane H(+)-ATPase of Candida albicans. |
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Authors: | P Gupta S K Mahanty S Ansari R Prasad |
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Institution: | School of Life Sciences, Jawaharlal Nehru University, New Delhi, India. |
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Abstract: | The plasma membrane ATPase of Candida albicans was solubilized by Tween 40 and purified to homogeneity on glycerol step gradient. The purified protein appeared as a single band of 100 +/- 4 KDa, represented greater than 98% of the total pure protein on densitometer scan. The purified PM-ATPase which was very specific to MgATP, had Km of about 0.77 mM and a sharp pH optimum at 6.6. Orthovanadate was able to inhibit the enzyme in a non-competitive manner, however, at higher concentrations the nature of inhibition changed to uncompetitive type. Based on molecular size, immuno cross-reactivity and sensitivity to different inhibitors, PM-ATPase of C. albicans appears to be similar to other ion pumps. |
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