Synthesis of fluorescent and radiolabeled analogues of phosphatidic acid.
Procedures for the synthesis of fluorescent and radiolabeled analogues of phosphatidic acid are described. The fluorophore 7-nitrobenzo-2-oxa-1,3-diazole (NBD) was coupled to 6-amino-caproic acid and 12-aminododecanoic acid by reaction of NBD-chloride with the amino acids under mild alkaline conditions at room temperature. 1,2-Dioleoyl-sn-[U-14C]glycerol 3-phosphate was prepared by acylation of sn-[U-14C]glycerol 3-phosphate with oleic acid anhydride using dimethylaminopyridine as the catalyst. This compound was converted to 1-oleoyl-sn-[U-14C]glycerol 3-phosphate by hydrolysis with phospholipase A2. The lysophosphatidic acid was reacylated with NBD-aminocaproyl imidazole or NBD-aminododecanoyl imidazole to form the fluorescent, radiolabeled analogue of phosphatidic acid. Fluorescent, non-radiolabeled analogues of phosphatidic acid were prepared by phospholipase D hydrolysis of fluorescent phosphatidylcholine.
D009327:4-Chloro-7-nitrobenzofurazan; D000215:Acylation; D015119:Aminocaproic Acid; D002250:Carbon Radioisotopes; D005456:Fluorescent Dyes; D007553:Isotope Labeling; D007850:Lauric Acids; D010712:Phosphatidic Acids / Q000138:chemical synthesis*
D002250:Carbon Radioisotopes; D005456:Fluorescent Dyes; D007850:Lauric Acids; D010712:Phosphatidic Acids; C046049:12-aminododecanoic acid; D009327:4-Chloro-7-nitrobenzofurazan; D015119:Aminocaproic Acid