Compared selectivities of the phosphatidylinositol-synthase from maize coleoptiles either in microsomal membranes or after solubilization.
PI-synthase selectivity from etiolated maize coleptiles was studied either associated with the microsomal membranes or after solubilization by CHAPS and prepurification on a DEAE-trisacryl M column. When maize microsomes were incubated with [3H]inositol without any exogenous CPM-PA, the most heavily labelled molecular species were 16:0/18:2-PI (77%), 16:0/18:3-plus 18:2/18:2-PI (15%), 16:0/18:1-PI (4%) and 18:0/18:2-PI (4%). Addition to the incubation medium of up to 300 microM 16:0/16:0-CMP-PA unexpectedly resulted in the formation of very little labelled 16:0/16:0-PI. When the solubilized fraction from microsomes was incubated with [3H]inositol in absence of 16:0/16:0-CPM-PA, the same PI molecular species as above were synthesized. However, with increasing concentrations of 16:0/16:0-CMP-PA in the medium, increasing amounts of labelled 16:0/16:0-PI appeared as well. With prepurified PI-synthase eluted from a DEAE column, endogenous CMP-PA was poorly utilized for PI biosynthesis whereas the exogenous 16:0/16:0-CPM-PA was used actively. With time, the endogenous CMP-PA was utilized first and the exogenous substrate was utilized, albeit, much more slowly. The results demonstrate that the selectivity displayed by PI-synthase towards various molecular species of CMP-PA depends on the integration of the enzyme in the membrane structure. Solubilization of the enzyme, i.e., inclusion of the protein in micelles with detergents and lipids, results in an apparent loss of the selectivity for CMP-PA.
Justin
A M
AM
Laboratoire de Physiologie Cellulaire et Moléculaire des Plantes, URA CNRS 1180, Université Pierre et Marie Curie, Paris, France.
Hmyene
A
A
Kader
J C
JC
Mazliak
P
P
D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; D002793:Cholic Acids; D008861:Microsomes / Q000201:enzymology; D012995:Solubility; D013347:Subcellular Fractions / Q000201:enzymology; D013379:Substrate Specificity; D017855:Transferases (Other Substituted Phosphate Groups) / Q000302:isolation & purification / Q000378:metabolism*; D003313:Zea mays / Q000196:embryology / Q000201:enzymology*
D002793:Cholic Acids; D017855:Transferases (Other Substituted Phosphate Groups); D051607:CDP-Diacylglycerol-Inositol 3-Phosphatidyltransferase; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate