Na(+)-dependent bile acid transport by hepatocytes is mediated by a protein similar to microsomal epoxide hydrolase.
A protein mediating hepatocyte sodium-dependent bile acid transport across the sinusoidal plasma membrane has been purified by immunoprecipitation with monoclonal antibody (MAb) 25D-1, which specifically recognizes this protein on the surface of intact hepatocytes (Ananthanarayanan et al. J. Biol. Chem. 263: 8338-8343, 1988). The function of this protein was further established by proteoliposome reconstitution (von Dippe et al. J. Biol. Chem. 265: 14812-14816, 1990). NH2-terminal amino acid sequence analysis and amino acid composition revealed this protein to be closely related to the enzyme microsomal epoxide hydrolase (mEH). Both proteins exhibited the same elution times on a reverse-phase high-pressure liquid chromatography column, comigrated with an apparent molecular weight of 49,000 as measured by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and possessed identical isoelectric points of 8.2. The MAb was capable of immunoprecipitating chromatographically purified mEH, as well as a protein derived from the sinusoidal plasma membrane that exhibited mEH activity comparable to that of the protein isolated from the endoplasmic reticulum. The subtilisin fragmentation patterns derived from chromatographically purified mEH and the MAb-precipitated plasma membrane protein were also identical. Hydropathy profile analysis of the amino acid sequence of mEH suggested the presence of four transmembrane domains. The results of these studies indicate that a protein that is involved in mediating sodium-dependent bile acid transport is closely related to mEH.
Von Dippe
P
P
Department of Biochemistry, School of Medicine, University of Southern California, Los Angeles 90033.
Amoui
M
M
Alves
C
C
Levy
D
D
D000818:Animals; D000911:Antibodies, Monoclonal; D001647:Bile Acids and Salts / Q000493:pharmacokinetics*; D001692:Biological Transport / Q000502:physiology; D002462:Cell Membrane / Q000502:physiology / Q000648:ultrastructure; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D004851:Epoxide Hydrolases / Q000032:analysis / Q000502:physiology*; D007525:Isoelectric Focusing; D008099:Liver / Q000166:cytology* / Q000502:physiology* / Q000648:ultrastructure; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology* / Q000502:physiology / Q000648:ultrastructure; D011233:Precipitin Tests; D051381:Rats; D017207:Rats, Sprague-Dawley; D012964:Sodium / Q000502:physiology*
D000911:Antibodies, Monoclonal; D001647:Bile Acids and Salts; D012964:Sodium; D004851:Epoxide Hydrolases