Hepatic microsomal glucuronidation of bilirubin is modulated by the lipid microenvironment of membrane-bound substrate.
Hepatocyte intracellular membranes may facilitate the directed movement of bilirubin and other hydrophobic substrates to the active site of UDP-glucuronyltransferase in the endoplasmic reticulum. We postulated that the lipid composition and physical properties of membranes that transport substrate may modulate bilirubin glucuronidation. To examine this hypothesis, we incorporated [14C]bilirubin substrate into the membrane bilayer of small unilamellar liposomes composed of native phospholipid purified from rat hepatic microsomes. The initial velocity of bilirubin glucuronide formation in rat liver microsomes, measured by radiochemical assay, was considerably more rapid than for bilirubin in liposomes of egg phosphatidylcholine (p less than 0.001). Moreover, the ratio of bilirubin diglucuronide to monoglucuronides synthesized was markedly increased (p less than 0.01), approaching that observed in normal rat bile. Although the rates of bilirubin glucuronidation did not correlate with fluidity of the liposomal membrane core region, specific phospholipid head groups were associated with an increase, and cholesterol a decrease, in rates of glucuronidation. Movement of [3H]bilirubin from dual-labeled liposomes to microsomes occurred without concomitant [14C]phospholipid transfer. Thus, the lipid composition of membranes incorporating bilirubin appears to modulate the rate of glucuronidation and the relative rates of bilirubin mono- and diglucuronide formation. Phospholipid head groups on the surface of the bilayer, not the hydrocarbon core regions, may be implicated in the rapid process of membrane transport, which is likely to involve membrane-membrane collisions or diffusion of free substrate rather than membrane fusion.
Whitmer
D I
DI
Russell
P E
PE
Ziurys
J C
JC
Gollan
J L
JL
D000818:Animals; D001663:Bilirubin / Q000031:analogs & derivatives* / Q000378:metabolism*; D002250:Carbon Radioisotopes; D002784:Cholesterol / Q000378:metabolism; D066298:In Vitro Techniques; D007700:Kinetics; D008027:Light; D008081:Liposomes / Q000378:metabolism; D008297:Male; D008560:Membrane Fluidity; D008563:Membrane Lipids / Q000378:metabolism*; D008862:Microsomes, Liver / Q000032:analysis / Q000201:enzymology*; D010743:Phospholipids / Q000032:analysis; D051381:Rats; D011919:Rats, Inbred Strains; D012542:Scattering, Radiation; D014316:Tritium
D002250:Carbon Radioisotopes; D008081:Liposomes; D008563:Membrane Lipids; D010743:Phospholipids; D014316:Tritium; C019270:bilirubin diglucuronide; C016266:bilirubin glucuronate; D002784:Cholesterol; D001663:Bilirubin