Effect of apoprotein B conformation on the activation of lysolecithin acyltransferase and lecithin: cholesterol acyltransferase. Studies with subfractions of low density lipoproteins.
In order to determine the role of apoprotein (apo) B conformation in the activation of the lysolecithin acyl-transferase reaction, we studied the activation of purified enzyme by various subfractions of low density lipoprotein (LDL), isolated by density gradient centrifugation. The activation of LAT correlated positively with the density of LDL and negatively with cholesterol/protein and triglyceride (TG)/protein ratios. The enzyme activation was also positively correlated with the number of trinitrobenzenesulfonic acid-reactive lysine amino groups, which increased with increasing density of LDL. The immunoaffinity of the LDL subfractions for B1B6, a monoclonal antibody directed to the receptor-binding region of apoB, increased with increasing density, while the affinity toward C1.4, a monoclonal antibody directed to the amino-terminal region of apoB, was not altered. Enrichment of normal whole LDL with TG resulted in a 45% reduction in enzyme activation, a 27% decrease in the number of trinitrobenzenesulfonic acid-reactive lysine groups, and a marked reduction in the immunoaffinity for B1B6. All these parameters reversed to normal when the TG-enriched LDL was treated with milk lipoprotein lipase, which specifically reduced the TG content of LDL. The LDL subfractions also supported cholesterol esterification by the purified enzyme, in parallel with lysolecithin esterification, indicating that apoB can also serve as an activator of the lecithin-cholesterol acyltransferase reaction. These results strongly suggest that the localized conformational change of apoB which occurs during the TG depletion of the precursor particle is critical for its activation of acyltransferase reactions, in a manner analogous to its interaction with the cellular receptors.
Liu
M
M
Department of Medicine, Rush Medical College, Chicago, Illinois 60612.
Krul
E S
ES
Subbaiah
P V
PV
D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000097:blood* / Q000302:isolation & purification; D001055:Apolipoproteins B / Q000737:chemistry / Q000494:pharmacology*; D002499:Centrifugation, Density Gradient; D002784:Cholesterol / Q000097:blood; D004789:Enzyme Activation; D006801:Humans; D007700:Kinetics; D008077:Lipoproteins, LDL / Q000097:blood / Q000302:isolation & purification / Q000494:pharmacology*; D007862:Phosphatidylcholine-Sterol O-Acyltransferase / Q000097:blood* / Q000302:isolation & purification; D010743:Phospholipids / Q000097:blood; D011487:Protein Conformation; D014280:Triglycerides / Q000097:blood / Q000494:pharmacology
D001055:Apolipoproteins B; D008077:Lipoproteins, LDL; D010743:Phospholipids; D014280:Triglycerides; D002784:Cholesterol; D008243:1-Acylglycerophosphocholine O-Acyltransferase; D007862:Phosphatidylcholine-Sterol O-Acyltransferase