Deacylation of bis(monoacylglycero)phosphate by lysosomal and microsomal lysophospholipases from rat liver.
The degradation of bis(monoacylglycero)phosphate by subcellular fractions of rat liver, using substrates labelled biosynthetically with [14C]oleic acid and chemically by catalytic exchange with tritium, was studied. Liver homogenates catalyzed maximum degradation at alkaline pH and subcellular fractionation localized this activity to microsomes. The degradation by microsomes was found to be a deacylation to lysophosphatidylglycerol and was without phosphodiesterase activity. The deacylation was maximal at pH 8.3 and did not require Ca2+ or Mg2+ but was stimulated by ethylenediaminetetraacetic acid and inhibited by Fe2+ and Hg2+. It was also inhibited by p-chloromercuribenzoate, deoxycholate, Triton X-100, and Triton WR-1339. The apparent Km was determined to be 5.5 X 10(-5) M and the corresponding V max was 4.1 nmol product released/min per milligram protein. The three labelled substrates were degraded by microsomes to give the same products in similar relative proportions. Degradation of bis(monoacylglycero)phosphate by lysosomes was maximal at acid pH as previously described by Y. Matsuzawa and K. Y. Hosteler. Contrary to their finding, deacylase activity in lysosomes was much greater than phosphodiesterase activity. The lysosomal deacylase but not the phosphodiesterase activity was inhibited reversibly by n-butanol. Sphingomyelin inhibited the microsomal deacylase but not the lysosomal deacylase.
D020001:1-Butanol; D000818:Animals; D016718:Arachidonic Acid; D001095:Arachidonic Acids / Q000378:metabolism; D000440:Butanols / Q000494:pharmacology; D005227:Fatty Acids / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D066298:In Vitro Techniques; D008245:Lysophospholipase / Q000378:metabolism*; D008246:Lysophospholipids; D008247:Lysosomes / Q000201:enzymology*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology*; D050178:Monoglycerides; D010712:Phosphatidic Acids / Q000378:metabolism*; D010740:Phospholipases / Q000378:metabolism*; D011506:Proteins / Q000378:metabolism; D051381:Rats; D013347:Subcellular Fractions / Q000378:metabolism
D001095:Arachidonic Acids; D000440:Butanols; D005227:Fatty Acids; D008246:Lysophospholipids; D050178:Monoglycerides; D010712:Phosphatidic Acids; D011506:Proteins; C012786:bis(monoacylglyceryl)phosphate; D016718:Arachidonic Acid; D020001:1-Butanol; D010740:Phospholipases; D008245:Lysophospholipase