Phosphatidylglycerol biosynthesis in Bacillus licheniformis Resolution of membrane-bound enzymes by affinity chromatography on cytidinediphospho-sn-1,2-diacylglycerol Sepharose.
Cytidinediphospho-sn-1,2-diaclglycerol (CDP-diglyceride) has been covalently linked to Sephrose 4B via adipic acid dihydrazide spacer arm forming an effective affinity chromatography column.  This liponucleo-tide ligand and sn-glycero-3-phosphate are subtracts for the formation of 3-sn-phoshatidyl-1'-sn-glycero-3'-phosphate (PGP) catalyzed in both eukaryotic and prokaryotic organisms by sn-glycero-3-phosphate:  CMP phosphatidlytranferase (PGP synthetase).  Using this CDP-diglyceride Sephrose affinity column we were able to resolve the membrane associated 3-sn-phosphatidyl'1-sn-glycerol (PG) synthesizing system present in Bacillus licheniformis into two activities.  A PGP synthetase activity was adsorbed to the affinity column and was eluted using buffer containg CDP-diglyceride; a PGP phosphatease acactivity had no affinity for the column.  Both PGP synthase and PGP phosphatase of B.  licheniformis were associated with a membrane component of the cell as evidenced by sucrose gradient centrifugation, differential centrifugation, and solubilization by buffers containing detergent...
D001407:Bacillus / Q000201:enzymology*; D002462:Cell Membrane / Q000187:drug effects / Q000201:enzymology; D002846:Chromatography, Affinity; D003567:Cytidine Diphosphate Diglycerides; D010715:Phosphatidylglycerols / Q000096:biosynthesis*; D010743:Phospholipids / Q000096:biosynthesis*; D010744:Phosphoric Monoester Hydrolases / Q000302:isolation & purification / Q000378:metabolism; D010770:Phosphotransferases / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D012685:Sepharose; D017855:Transferases (Other Substituted Phosphate Groups)
D003567:Cytidine Diphosphate Diglycerides; D010715:Phosphatidylglycerols; D010743:Phospholipids; D011092:Polyethylene Glycols; D012685:Sepharose; D010770:Phosphotransferases; D017855:Transferases (Other Substituted Phosphate Groups); C020676:CDP-diacylglycerol-glycerol-3-phosphate 3-phosphatidyltransferase; D010744:Phosphoric Monoester Hydrolases