Kinetics of bilirubin transfer between serum albumin and membrane vesicles. Insight into the mechanism of organic anion delivery to the hepatocyte plasma membrane.
Unconjugated bilirubin is transported in the plasma bound primarily to serum albumin, from which it is taken up and metabolized by the liver. To better characterize the mechanism of bilirubin delivery to the hepatocyte, stopped-flow techniques were utilized to study the kinetics of bilirubin transfer between serum albumin and both model phospholipid and native hepatocyte plasma membrane vesicles. The transfer process was best described by a single exponential function, with rate constants of 0.93 +/- 0.04, 0.61 +/- 0.03, and 0.10 +/- 0.01 s-1 (+/- S.D.) at 25 degrees C for human, rat, and bovine serum albumins, respectively. The observed variations in rate with respect to donor and acceptor concentrations provide strong evidence for the diffusional transfer of free bilirubin. Thermodynamic analysis suggests that the binding site on bovine serum albumin demonstrates higher specificity for the bilirubin molecule than that on human or rat serum albumin, which exhibit similar binding characteristics. Kinetic analysis of bilirubin transfer from rat serum albumin to isolated rat basolateral liver plasma membranes indicates that the delivery of albumin-bound bilirubin to the hepatocyte surface occurs via aqueous diffusion, rather than a collisional process, thereby mitigating against the presence of an "albumin receptor."
Division of Gastroenterology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
S D
SD
Zucker
W
W
Goessling
J L
JL
Gollan
D000818:Animals; D001663:Bilirubin / Q000737:chemistry / Q000378:metabolism*; D001692:Biological Transport; D002417:Cattle; D006801:Humans; D066298:In Vitro Techniques; D017136:Ion Transport; D007700:Kinetics; D008099:Liver / Q000166:cytology / Q000378:metabolism*; D008297:Male; D010713:Phosphatidylcholines / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D012709:Serum Albumin / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence; D013816:Thermodynamics
D010713:Phosphatidylcholines; D012709:Serum Albumin; D001663:Bilirubin