Regulation of microsomal enzymes by phospholipids. IX. Production of uniquely modified forms of microsomal UDP-glucuronyltransferase by treatment with phospholipase A and detergents.
The kinetic parameters of microsomal UDPglucuronyltransferase (EC 2.4.1.17) were compared after treatment of microsomes with phospholipase A and Triton X-100. Treatments with phospholipase A and Triton had differential effects on activities at V (assayed in both forward and reverse directions), the affinity of the enzyme for UDPglucuronic acid and UDP, the specificity of binding of UDPsugars, and the sensitivity of the enzyme to treatment with UDP-N-acetylgulcosamine. Comparisons of lysophosphatide, Triton, and cholate-activated forms indicated that different types of detergents activate by separate mechanisms. These data are taken as evidence that selective types of changes of the composition and structure of microsomal lipids lead to selective types of changes of the properties of UDPglucuronyltransferase. Activation of UDPglucuronyltransferase in response to different types of perturbations of the membrane lipids is thus a specific process. It was found that constraint on the maximal potential activity of the reverse reaction catalyzed by UDPglucuronyltransferase is more extensive in untreated microsomes than constraint on the forward reaction. Also, the "activated forms" of UDPglucuronyltransferase have less activity than the untreated enzyme if assays are carried out under conditions presumed to exist in vivo. These results indicate the complexity of lipid-protein interactions as regulators of membrane-bound enzymes. They also indicate the functional significance of this type of regulation for the activity of UDPglucuronyltransferase.
D000818:Animals; D001665:Binding Sites; D002793:Cholic Acids / Q000494:pharmacology; D014453:Glucuronosyltransferase / Q000378:metabolism*; D006168:Guinea Pigs; D006602:Hexosyltransferases / Q000378:metabolism*; D007700:Kinetics; D008244:Lysophosphatidylcholines / Q000494:pharmacology; D008862:Microsomes, Liver / Q000187:drug effects / Q000201:enzymology*; D010740:Phospholipases / Q000494:pharmacology*; D010743:Phospholipids / Q000494:pharmacology*; D011092:Polyethylene Glycols / Q000494:pharmacology*; D011485:Protein Binding; D000644:Quaternary Ammonium Compounds / Q000494:pharmacology*
D002793:Cholic Acids; D008244:Lysophosphatidylcholines; D010743:Phospholipids; D000644:Quaternary Ammonium Compounds; D011092:Polyethylene Glycols; D006602:Hexosyltransferases; D014453:Glucuronosyltransferase; D010740:Phospholipases