title: Isolation, purification and kinetic characterization of plasma membrane H(+)-ATPase of Candida albicans.
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.
authors.affiliation: School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.
authors.forename: P
authors.initials: P
authors.lastname: Gupta
authors.forename: S K
authors.initials: SK
authors.lastname: Mahanty
authors.forename: S
authors.initials: S
authors.lastname: Ansari
authors.forename: R
authors.initials: R
authors.lastname: Prasad
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D000906:Antibodies / Q000276:immunology; D002176:Candida albicans / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D003720:Densitometry; D004797:Enzyme-Linked Immunosorbent Assay; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D006180:Proton-Translocating ATPases / Q000302:isolation & purification / Q000378:metabolism*; D014638:Vanadates / Q000494:pharmacology
chemical_list: D000906:Antibodies; D014638:Vanadates; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases