Glycerolipid metabolizing enzymes in rat ventricle and in cardiac myocytes.
1. The properties and subcellular distribution of phosphatidate phosphohydrolase (PAP) were studied in rat heart. A Mg2(+)-activated activity (PAP1) which was inhibited by N-ethylmaleimide was found mainly in a 105,000 x g soluble fraction. Isolation of the membranes in a medium containing KCl increased the proportion of PAP1 that was associated. Translocation of PAP1 from these membranes occurred on subsequent incubation in a low-ionic strength medium from which KCI was omitted. Incubation of cardiac myocytes with palmitate promoted translocation of PAP activity to cellular membranes. A second activity which was insensitive to N-ethylmaleimide (PAP2) was found in the 105,000 x g membrane fraction. PAP2 was inhibited by concentrations of Mg2+ known to occur in ischaemia. Specific activities of PAP1 and PAP2 in ventricle muscle homogenates were similar. The specific activity of PAP2 in homogenates of cardiac myocytes was only 42% of that in homogenates of ventricle muscle. 2. A glycerolphosphate acyltransferase (GPAT) activity with properties similar to the GPAT found in microsomes from liver or adipose tissue was enriched in the sarcoplasmic reticulum fraction from ventricle muscle. This GPAT had a significantly higher K(m) for glycerol 3-phosphate than the GPAT found in adipose tissue microsomes. The possible physiological significance of this 'high K(m)' GPAT in heart, particularly in ischaemia, is discussed. 3. Comparisons were made of the specific activities of fatty acyl-CoA synthetase, monoacylglycerolphosphate acyltransferase, diacylglycerol acyltransferase and the mitochondrial and microsomal forms of GPAT in homogenates from cardiac myocytes and ventricle muscle.
Swanton
E M
EM
Department of Biochemistry and Molecular Biology, University College London, UK.
Saggerson
E D
ED
D000818:Animals; D005992:Glycerol-3-Phosphate O-Acyltransferase / Q000378:metabolism*; D006352:Heart Ventricles; D066298:In Vitro Techniques; D008297:Male; D009206:Myocardium / Q000166:cytology / Q000201:enzymology*; D000073718:Pancreatitis-Associated Proteins; D010711:Phosphatidate Phosphatase / Q000037:antagonists & inhibitors / Q000378:metabolism*; D051381:Rats; D017207:Rats, Sprague-Dawley; D013347:Subcellular Fractions / Q000201:enzymology
D000073718:Pancreatitis-Associated Proteins; C000612141:REG3A protein, human; D005992:Glycerol-3-Phosphate O-Acyltransferase; D010711:Phosphatidate Phosphatase