Phosphatidylinositol synthesis and exchange of the inositol head are catalysed by the single phosphatidylinositol synthase 1 from Arabidopsis.
In order to study some of its enzymatic properties, phosphatidylinositol synthase 1 (AtPIS1) from the plant Arabidopsis thaliana was expressed in Escherichia coli, a host naturally devoid of phosphatidylinositol (PtdIns). In the context of the bacterial membrane and in addition to de novo synthesis, the plant enzyme is capable of catalysing the exchange of the inositol polar head for another inositol. Our data clearly show that the CDP-diacylglycerol-independent exchange reaction can occur using endogenous PtdIns molecular species or PtdIns molecular species from soybean added exogenously. Exchange has been observed in the absence of cytidine monophosphate (CMP), but is greatly enhanced in the presence of 4 microm CMP. Our data also show that AtPIS1 catalyses the removal of the polar head in the presence of much higher concentrations of CMP, in a manner that suggests a reverse of synthesis. All of the PtdIns metabolizing activities require free manganese ions. EDTA, in the presence of low Mn2+ concentrations, also has an enhancing effect.
Justin
Anne-Marie
AM
Université Pierre et Marie Curie and CNRS, Laboratoire de Physiologie Cellulaire et Moléculaire, Paris, France.
Kader
Jean-Claude
JC
Collin
Sylvie
S
D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000201:enzymology*; D029681:Arabidopsis Proteins; D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; D002384:Catalysis; D004492:Edetic Acid / Q000494:pharmacology; D008345:Manganese / Q000494:pharmacology; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000096:biosynthesis*; D017855:Transferases (Other Substituted Phosphate Groups) / Q000378:metabolism*
D029681:Arabidopsis Proteins; D008565:Membrane Proteins; D010716:Phosphatidylinositols; D008345:Manganese; D004492:Edetic Acid; D017855:Transferases (Other Substituted Phosphate Groups); C496769:ATPIS1 protein, Arabidopsis; D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase