Identification of the hepatocyte sodium-dependent bile acid transport protein using a monoclonal antibody

Conference · Fri May 01 04:00:00 EDT 1987 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:5856686

Levy, D; Ananthanarayanan, M; von Dippe, P

Hepatocytes have been shown to possess a sodium-dependent and sodium-independent bile acid transport system. Photoaffinity labeling studies have identified the carrier proteins as two intrinsic membrane components (54 kDa and 49 kDa) located on the sinusoidal surface domain. These proteins were also labeled by DIDS which is a potent inhibitor of hepatocyte bile acid transport. These two proteins have been significantly co-purified by affinity chromatography and subsequently utilized as an immunogen for the production of monoclonal antibodies. Spleen cells from immunized SJL-J mice were fused with SP2/0 Ag 14 myeloma cells and the resulting hybridoma colonies screened by ELISA and immunoprecipitation assays. Colonies producing antibodies against these proteins were cloned and IgG mabs purified from ascites fluid on a protein A-Sepharose column. Several mabs were shown to bind to a 49 kDa protein on the surface of intact hepatocytes, which was one of the proteins labeled by DIDS. The binding of one of these antibodies (25A3) to hepatocytes resulted in a substantial protection of the Na/sup +/-dependent bile acid transport system from inhibition by DIDS. Binding studies with /sup 125/I-25A3 indicated that this bile acid carrier comprised approximately 1% of the total membrane protein. These results establish that a 49 kDa protein in hepatocyte sinusoidal plasma membranes is a component of the sodium-dependent bile acid transport system.

• Research Organization: Univ. of Southern California, Los Angeles
• OSTI ID: 5856686
• Report Number(s): CONF-870644-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 46:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ALKALI METAL COMPOUNDS
ANIMAL CELLS
ANIMALS
ANTIBODIES
BETA DECAY RADIOISOTOPES
BILE ACIDS
BIOCHEMICAL REACTION KINETICS
CARBOXYLIC ACIDS
CATIONS
CHARGED PARTICLES
CLONING
DAYS LIVING RADIOISOTOPES
ELECTRON CAPTURE RADIOISOTOPES
HYBRIDOMAS
HYDROXY COMPOUNDS
INTERMEDIATE MASS NUCLEI
IODINE 125
IODINE ISOTOPES
IONS
ISOTOPE APPLICATIONS
ISOTOPES
KINETICS
LABELLED COMPOUNDS
LIVER CELLS
MAMMALS
MEMBRANE TRANSPORT
MICE
MOLECULAR WEIGHT
MONOCLONAL ANTIBODIES
NUCLEI
ODD-EVEN NUCLEI
ORGANIC ACIDS
ORGANIC COMPOUNDS
PROTEINS
PURIFICATION
RADIOISOTOPES
REACTION KINETICS
RODENTS
SODIUM COMPOUNDS
SOMATIC CELLS
SPLEEN CELLS
STEROIDS
STEROLS
TRACER TECHNIQUES
TUMOR CELLS
VERTEBRATES