Detergent properties of water-soluble choline phosphatides. Selective solubilization of acyl-CoA:lysolecithin acyltransferase from thymocyte plasma membranes.
Several analogs of lysolecithin were found to solubilize human erythrocyte ghosts comparably or even better than other detergents. Derivatives with aliphatic chains of 12 to 14 carbons were most effective. The phosphorylcholine detergents apparently possess low protein-denaturing properties, since they, for the first time, allowed the solubilization of enzymatically active acyl-CoA:lysolecithin acyltransferase from thymocyte plasma membranes. The solubilized enzyme was not sedimented at 177,000 x g for 60 min and penetrated into Sepharose 6B gels. Low detergent concentration resulted in a selective extraction of the acyltransferase (about 70%) as compared to alkaline phosphatase, nucleotide pyrophosphatase, gamma-glutamyltransferase or Mg2+-ATPase (30 to 40%). The selectivity was reflected in sodium dodecyl sulfate-polyacrylamide gel electrophoresis patterns of soluble and sedimentable membrane fractions; three bands of approximately 53, 84, and 94 x 10(3) daltons were enriched in the supernatants, whereas one band of about 68 x 10(3) daltons was concentrated in the pellet. The preferential extraction of acyltransferase may be related to particularly high affinity of lysolecithin analogs for this enzyme, which at higher concentrations was competitively inhibited by these detergents. The inhibitor constants ranged from 1400 micron for the C10 analog (ET-10-H) to 80 micron for the compound with 16 carbons (ET-16-H) per aliphatic chain.
D000818:Animals; D002417:Cattle; D002462:Cell Membrane / Q000201:enzymology; D004910:Erythrocyte Membrane / Q000032:analysis; D006801:Humans; D007700:Kinetics; D008563:Membrane Lipids / Q000097:blood / Q000494:pharmacology; D008970:Molecular Weight; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000302:isolation & purification*; D010713:Phosphatidylcholines / Q000494:pharmacology; D010743:Phospholipids / Q000097:blood / Q000494:pharmacology; D012995:Solubility; D013950:Thymus Gland / Q000201:enzymology*
D008563:Membrane Lipids; D010713:Phosphatidylcholines; D010743:Phospholipids; D007862:Phosphatidylcholine-Sterol O-Acyltransferase