Characteristics of mitochondrial and microsonal monoacyl- and diacylglycerol 3-phosphate biosynthesis in rabbit heart.

Abstract

Acylation of sn-glycerol 3-phosphate by heart subcellular fractions was characterized. The enzyme kinetics revealed that the rate of reaction of acylation by mitochondria was slower, but constant for a longer period (up to 20min), than that by the microsomal fraction. The range of palmitate, oleate and linoleate concentrations yielding optimal sn-glycerol 3-phosphate acylation was broader for mitochondria than for the microsomal fraction, the latter showing a preference for linoleate. The mitochondrial fraction synthesized a relatively large quantity of monoacyl-sn-glycerol 3-phosphate, reaching 135% of the microsomal biosynthesis during an assay period of 15min. By contrast, the microsomal fraction formed considerably more diacyl- than monoacyl-sn-glycerol 3-phosphate, except with linoleate as the acyl donor, in which case approximately equal quantities of the two products were produced. The biosynthesis of monoacyl-sn-glycerol 3-phosphate was also observed in experiments in which hepatic subcellular fractions were used to provide supporting evidence. Cardiac mitochondrial diacyl-sn-glycerol 3-phosphate formation was less than 17% of the microsomal formation. However, evidence is presented to exclude the possibility that monoacyl-sn-glycerol 3-phosphate in the mitochondrial fraction is formed by deacylation of the contaminating microsomal diacyl-sn-glycerol 3-phosphate. The participation of the dihydroxyacetone phosphate pathway in the biosynthesis of these substances was minimal. The addition of CTP and the fatty acid specificity of the reaction both provided results that reinforced the postulate that mitochondrial differs from microsomal acylation. Thus our findings demonstrate that the characteristics of acyl-CoA-sn-glycerol 3-phosphate O-acyltransferase (EC 2.3.1.15) in rabbit heart mitochondria are distinct from those of cardiac microsomal enzyme and hepatic enzymes.