Candida albicans phosphatidylinositol synthase has common features with both Saccharomyces cerevisiae and mammalian phosphatidylinositol synthases.
Phosphatidylinositol (PI) synthase (cytidine 5'-diphospho (CDP)-1,2-diacyl-sn-glycerol:myo-inositol 3-phosphatidyltransferase, EC 2.7.8.11) was isolated from the microsomal cell fraction of Candida albicans. The Triton X-100 extracted enzyme was enriched 140-fold by affinity chromatography on CDP-diacylglycerol-Sepharose. The enzyme had a pH optimum at 9.5 in glycine/NaOH buffer. It had an absolute requirement for Mg2+ or Mn2+ and was inhibited by Ca2+ and Zn2+. Maximal activity was at 0.2-0.6 mM-CDP-diacylglycerol, higher concentrations inhibited the enzyme. With 2'-deoxy-CDP-diacylglycerol as the lipid substrate, optimal activity was at 0.7 mM. The K(m) for myo-inositol was determined to be 0.55 mM. The optimal temperature for the PI synthase reaction was 55 degrees C. The C. albicans PI synthase shows differences to the Saccharomyces cerevisiae enzyme, such as activation by bivalent cations, inhibition by nucleotides, temperature optimum and activation energy, but also to the human PI synthase in preference for the lipid substrates, inhibition by nucleoside monophosphates and stabilization by Mn2+ and phospholipids.
Antonsson
B E
BE
Glaxo Institute for Molecular Biology, Geneva, Switzerland.
Klig
L S
LS
D000818:Animals; D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; D002176:Candida albicans / Q000201:enzymology*; D002712:Chlorides / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015636:Magnesium Chloride / Q000494:pharmacology; D008322:Mammals; D017895:Manganese Compounds / Q000494:pharmacology; D009711:Nucleotides / Q000494:pharmacology; D010710:Phosphates / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D013379:Substrate Specificity; D013696:Temperature; D017855:Transferases (Other Substituted Phosphate Groups) / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*
D002712:Chlorides; D004791:Enzyme Inhibitors; D017895:Manganese Compounds; D009711:Nucleotides; D010710:Phosphates; D015636:Magnesium Chloride; D017855:Transferases (Other Substituted Phosphate Groups); D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; C025340:manganese chloride