Comparison of the HPLC-separated species patterns of phosphatidic acid, CDP-diacylglycerol and diacylglycerol synthesized de novo in rat liver microsomes (a new method).
The species pattern of phosphatidic acid was compared with that of CDP-diacylglycerol and diacylglycerol synthesized de novo by glycerol 3-phosphate acylation in a CoA ester-generating system in liver microsomes. The similarity of the species patterns of phosphatidic acid and CDP-diacylglycerol indicated that the CTP-phosphatidyl cytidylyltransferase showed no selectivity for individual species of its phosphatidic acid substrate. Since the species pattern of diacylglycerol deviated from that of phosphatidic acid, a slight acyl selectivity of the phosphatidic acid phosphohydrolase or a slight inhomogeneity of its substrate pool might be assumed. For the determination of the molecular species of CDP-diacylglycerol, a new method was developed. By incubation of CDP-diacylglycerol with oligonucleate 5'-nucleotidohydrolase (phosphodiesterase), phosphatidic acid was produced. The CDP-diacylglycerol-derived phosphatidic acid was methylated with diazomethane and then separated by reverse-phase HPLC in 15 molecular species.
Rüstow
B
B
Institute of Pathological and Clinical Biochemistry, Charite Hospital, Humboldt University, Berlin, D.D.R.
Nakagawa
Y
Y
Rabe
H
H
Reichmann
G
G
Kunze
D
D
Waku
K
K
D000818:Animals; D002851:Chromatography, High Pressure Liquid; D003567:Cytidine Diphosphate Diglycerides / Q000302:isolation & purification*; D004075:Diglycerides / Q000302:isolation & purification*; D005989:Glycerides / Q000302:isolation & purification*; D008297:Male; D008722:Methods; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism*; D009702:Nucleoside Diphosphate Sugars / Q000302:isolation & purification*; D009713:Nucleotidyltransferases / Q000378:metabolism; D010711:Phosphatidate Phosphatase / Q000378:metabolism; D010712:Phosphatidic Acids / Q000302:isolation & purification*; D051381:Rats
D003567:Cytidine Diphosphate Diglycerides; D004075:Diglycerides; D005989:Glycerides; D009702:Nucleoside Diphosphate Sugars; D010712:Phosphatidic Acids; D009713:Nucleotidyltransferases; C022348:phosphatidate cytidylyltransferase; D010711:Phosphatidate Phosphatase