Purified arginine permease of Candida albicans is functionally active in a reconstituted system.
We have for the first time purified arginine permease from a pathogenic yeast, Candida albicans, to homogeneity by affinity chromatography using L-arginine-linked agarose matrix as affinity column. The purified protein (PP) was of 66 kDa with no subunit structure. Two kinetically distinct binding affinities of PP were evident where high affinity binding (S1) revealed a dependence on acidic pH while pH did not have dramatic effect on low affinity (S2) binding. The specificity of L-arginine binding to PP with regard to other amino acids, structural analogues and inhibitors, was essentially similar to arginine transport observed in the intact cells of C. albicans (Rao et al., 1986). The purified arginine permease was reconstituted into proteoliposomes and its functionality was tested by imposing a valinomycin-induced membrane potential. All the characteristic features of L-arginine transport displayed by the reconstituted system were similar to those observed in intact cells. Thus homogeneous purified arginine permease was also functionally active.
Mukherjee
P K
PK
School of Life Sciences, Jawaharlal Nehru University, New Delhi, India.
Prasad
R
R
D026905:Amino Acid Transport Systems; D026923:Amino Acid Transport Systems, Basic; D001120:Arginine / Q000378:metabolism; D001667:Binding, Competitive; D001692:Biological Transport; D002176:Candida albicans / Q000201:enzymology*; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D008081:Liposomes; D026901:Membrane Transport Proteins / Q000302:isolation & purification* / Q000378:metabolism*; D011485:Protein Binding; D011510:Proteolipids / Q000378:metabolism; D012995:Solubility; D013379:Substrate Specificity
D026905:Amino Acid Transport Systems; D026923:Amino Acid Transport Systems, Basic; D008081:Liposomes; D026901:Membrane Transport Proteins; D011510:Proteolipids; C020485:proteoliposomes; C043909:arginine permease; D001120:Arginine