The kinetics and divalent cation inhibition of plasma membrane ATPase in the yeast Candida albicans.
The kinetics of ATP hydrolysis and cation effects on ATPase activity in plasma membrane from Candida albicans ATCC 10261 yeast cells were investigated. The ATPase showed classical Michaelis-Menten kinetics for the hydrolysis of Mg X ATP, with Km = 4.8 mM Mg X ATP. Na+ and K+ stimulated the ATPase slightly (9% at 20 mM). Divalent cations in combination with ATP gave lower ATPase activity than Mg X ATP (Mg greater than Mn greater than Co greater than Zn greater than Ni greater than Ca). Divalent cations inhibited the Mg X ATPase (Zn greater than Ni greater than Co greater than Ca greater than Mn). Free Mg2+ inhibited Mg X ATPase weakly (20% inhibition at 10 mM). Computed analyses of substrate concentrations showed that free Zn2+ inhibited Zn X ATPase, mixed (Zn2+ + Mg2+) X ATPase, and Mg X ATPase activities. Zn X ATP showed high affinity for ATPase (Km = 1.0 mM Zn X ATP) but lower turnover (52%) relative to Mg X ATP. Inhibition of Mg X ATPase by (free) Zn2+ was noncompetitive, Ki = 90 microM Zn2+. The existence of a divalent cation inhibitory site on the plasma membrane Mg X ATPase is proposed.
Hubbard
M J
MJ
Sullivan
P A
PA
Shepherd
M G
MG
D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D002176:Candida albicans / Q000201:enzymology*; D002462:Cell Membrane / Q000201:enzymology; D007700:Kinetics; D011188:Potassium / Q000378:metabolism; D012964:Sodium / Q000378:metabolism; D015032:Zinc / Q000378:metabolism
D000255:Adenosine Triphosphate; D012964:Sodium; D000251:Adenosine Triphosphatases; D015032:Zinc; D011188:Potassium