Effect of lysolecithin in solubilizing membrane-bound glycosyltransferases.
Microsomal membranes were solubilized by incubation with lysolecithin which caused considerable release of galactosyl- and N-acetylglucosaminyl-transferase into a high-speed supernatant fraction. With a critical concentration of lysolecithin (2.5 mg/10 mg protein in 1 mL microsome suspension), there was a maximal binding of radioactive lysolecithin to the sediment fraction obtained after high-speed centrifugation. Increase of lysolecithin concentration (above 2.5 mg/mL) in the incubation mixture caused a progressive release of the enzymes into the supernatant fraction. Lysolecithin binding to the membrane was greatly inhibited by 1 M NaCl, and high salt concentration also inactivated galactosyltransferase in the sediment, suggesting an electrostatic interaction between lysolecithin and membrane enzyme. In contrast, high NaCl concentration had no inhibitory effect on the enzyme activity in the sediment when the fraction was prepared by treatment with Triton X-100. Lysolecithin-treated microsomal sediment and supernatant galactosyltransferase was inactivated by oleoyllysophosphatidic acid but not by palmitoyllysophosphatidic acid or egg yold lysophosphatidic acid. Triton X-100 treated microsomal fractions were also similarly affected by different species of lysophosphatidic acid. The results suggested a similarity of interactions of lysophosphatidic fatty acyl species with lysolecithin and Triton-treated galactosyltransferase.
D000818:Animals; D005700:Galactosyltransferases / Q000302:isolation & purification / Q000378:metabolism; D006027:Glycosides; D007700:Kinetics; D008244:Lysophosphatidylcholines; D008345:Manganese / Q000494:pharmacology; D008862:Microsomes, Liver / Q000201:enzymology*; D009994:Osmolar Concentration; D010743:Phospholipids / Q000494:pharmacology; D011092:Polyethylene Glycols; D051381:Rats; D014166:Transferases / Q000302:isolation & purification*
D006027:Glycosides; D008244:Lysophosphatidylcholines; D010743:Phospholipids; D011092:Polyethylene Glycols; D008345:Manganese; D014166:Transferases; D005700:Galactosyltransferases