Insertion of the hepatocyte bile acid carrier system into heaptoma tissue culture cells by membrane-cell fusion

Conference · Thu May 01 04:00:00 EDT 1986 · Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States)

OSTI ID:6899004

Levy, D; von Dippe, P

Hepatocytes have been shown to have a sodium-dependent bile acid transport system. The carrier protein for these substrates has been identified as a 54,000 dalton intrinsic membrane component. In an effort to reconstitute this transport system, studies have been concerned with the insertion of purified hepatocyte sinusoidal plasma membranes containing this carrier system, into the membranes of non-transporting cells by polyethylene glycol (PEG) mediated fusion procedures. Hepatoma tissue culture (HTC) cells were used in these studies since there is no detectable level of bile acid transport activity in this cell system. HTC cells were treated with 50% PEG in the presence of plasma membranes, while cells treated with either 50% PEG or plasma membranes alone served as controls. The resulting cells were then evaluated for transport capacity by fluorescence microscopy using a fluorescent 7-dansylhydrazone derivative of cholic acid, or by measurement of (/sup 3/H)taurocholic acid uptake. The transport of substrates by this reconstituted system was dependent on time, membrane protein concentration and a sodium gradient and could be inhibited by taurochenodeoxycholic acid. The control cells showed no measurable uptake of these substrates. These results indicate that this membrane fusion procedure will be useful in functional studies of this anion transport system.

• Research Organization: Univ. of Southern California, Los Angeles
• OSTI ID: 6899004
• Report Number(s): CONF-8606151-
• Conference Information: Journal Name: Fed. Proc., Fed. Am. Soc. Exp. Biol.; (United States) Journal Volume: 45:6
• Country of Publication: United States
• Language: English

Related Subjects

550201* -- Biochemistry-- Tracer Techniques
59 BASIC BIOLOGICAL SCIENCES
ANIMAL CELLS
BILE ACIDS
CARBOXYLIC ACIDS
CELL CONSTITUENTS
CELL MEMBRANES
DISEASES
FLUORESCENCE
HEPATOMAS
HYDROXY COMPOUNDS
ISOTOPE APPLICATIONS
LABELLED COMPOUNDS
LIVER CELLS
LUMINESCENCE
MEMBRANE PORES
MEMBRANE TRANSPORT
MEMBRANES
NEOPLASMS
ORGANIC ACIDS
ORGANIC COMPOUNDS
SOMATIC CELLS
STEROIDS
STEROLS
TISSUE CULTURES
TRACER TECHNIQUES
TRITIUM COMPOUNDS