Purification and reconstitution of the bile acid transport system from hepatocyte sinusoidal plasma membranes

Dippe, P von; Ananthanarayanan, M.; Drain, Peter; Lévy, Daniel

doi:10.1016/0005-2736(86)90238-5

Cited by 52 publications

(25 citation statements)

References 30 publications

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“…Affinity Column Purification of the Ileal 100-kDa Protein-The I100 protein was initially isolated based upon its ability to bind to an anionic glycocholate-Sepharose 4B affinity column (7). Ileal brush border membrane vesicles (BBMV) 1 were prepared from rat ileum by divalent cation precipitation as described previously (10).…”

Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of a Novel Peptidase from Rat and Human Ileum

Shneider

Thevananther

Moyer

et al. 1997

Journal of Biological Chemistry

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A novel 100-kDa ileal brush border membrane protein (I100) has been purified by anionic glycocholate affinity chromatography. Polyclonal antibodies raised against this protein were utilized to clone and characterize I100 in rats. A partial length human I100 cDNA was identified by hybridization screening. In the rat, the I100 protein is a 746-amino acid glycosylated (calculated core molecular mass of 80 kDa) type II integral membrane protein found on the apical surface of ileal villus enterocytes. Its 2.6-kilobase mRNA is expressed in distal small intestine in rats and in humans. The I100 cDNA is homologous to but distinct from human prostate-specific membrane antigen and rat brain N-acetylaspartylglutamate peptidase. It is expressed on both the basolateral and apical surfaces of stably transfected Madin Darby canine kidney cells. Analysis of these stably transfected Madin Darby canine kidney cells and I100 immunoprecipitates of rat ileal brush border membrane vesicles reveals that it has dipeptidyl peptidase IV activity. Future invesitgations will need to determine the exact substrate specificity of this novel peptidase.

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Section: Methodsmentioning

confidence: 99%

Cloning and Characterization of a Novel Peptidase from Rat and Human Ileum

Shneider

Thevananther

Moyer

et al. 1997

Journal of Biological Chemistry

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“…Thus, better preservation of the lipid environment and/or formation of more uniform and larger PLP may partially explain the better reconstitution of Na+ gradient-dependent taurocholate uptake into basolateral PLP (Fig. 5) compared to Triton X-100-solubilized sinusoidal plasma membrane proteins incorporated into liposomes prepared by sonication and freeze-thaw procedures (8).…”

Section: Discussionmentioning

confidence: 99%

“…The adopted reconstitution method was modeled after those previously described for other purposes (14)(15)(16). In principle, it consists of forming PLP directly from proteincontaining, phospholipid-supplemented, and detergent-rich mixed micelles (14) rather than starting with mixtures of phospholipid and protein in separate micellar solutions (8,28). By this means possible harmful delipidation and/or aggregation of hydrophobic proteins can be prevented.…”

Section: Discussionmentioning

confidence: 99%

“…The involved transport systems have recently been identified and partially characterized by using photoaffinity-labeling techniques and kinetic transport studies in intact hepatocytes as well as isolated plasma membrane vesicles (3)(4)(5)(6). Thus, basolateral uptake of bile salts is mediated by one or two polypeptides with molecular masses between 48 and 54 kDa (3,7,8). In contrast, a distinct polypeptide with a molecular mass of 100 kDa appears to be involved in the canalicular secretion of bile salts.…”

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confidence: 99%

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Functional reconstitution of the canalicular bile salt transport system of rat liver.

Ruetz

Hugentobler

Meier

1988

Proc. Natl. Acad. Sci. U.S.A.

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Recent studies have suggested that the canalicular bile salt transport system of rat liver corresponds to a 100-kDa membrane glycoprotein. In the present study we attempted to functionally reconstitute the 100-kDa protein into artificial proteoliposomes. Canalicular membrane proteins were solubilized with octyl glucoside in the presence ofasolectin phospholipids. The extracts were treated with preimmune serum or the 100-kDa protein selectively immunoprecipitated with a polyclonal antiserum. Proteins remaining in the supernatant were then incorporated into proteoliposomes by gel-filtration chromatography. Canalicular proteoliposomes containing the 100-kDa protein exhibited t ulatable taurocholate uptake that could be inhibited by 4,4'-diisothiocyanato-2,2'-stilbenedisulfonic acid (DmDS). In contrast, no DIDS-sensitive t imulatable taurocholate uptake was found in 100-kDa protein-free canalicular proteoliposomes. However, when the immunoprecipitated 100-kDa protein was dissociated from the antibodies and exclusively incorporated into liposomes, reconstitution of DIDSsensitive transstimulatable and electrogenic taurocholate anion transport was again positive. Although incorporation of solubilized basolateral membrane proteins into liposomes also resulted in a prompt reconstitution of Na gradient-driven taurocholate uptake, the anti-100-kDa antibodies had no effects on the reconstituted transport activity of basolateral proteins. Thus, the findings establish that the previously characterized canalicularspecific 100-kDa protein is directly involved in the transcanalicular secretion of bile salts.

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“…Several findings support this latter possibility. It was postulated that mEH binds to Na + -independent organic anion transport protein forming a multiple transport system [24]. Furthermore, mEH was reported to be associated with various cytochrome P450s [25,26], and also with phospholipids [27].…”

Section: Discussionmentioning

confidence: 99%

Autoantibody response to microsomal epoxide hydrolase in hepatitis C and A

Akatsuka

Kobayashi

Ishikawa

et al. 2007

Journal of Autoimmunity

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Autoimmune responses were observed in a large proportion of hepatitis C cases and are suspected to be part of viral pathogenesis. The AN6520 antigen (AN-Ag) is a normal cellular protein mainly expressed in liver that was found associated with non-A, non-B hepatitis. To elucidate its pathogenic role in hepatitis C, we developed an IgM capture assay using purified AN-Ag and confirmed that the antibody response to AN-Ag is associated almost exclusively with hepatitis C cases (29%). Screening of a human liver expression library revealed that AN-Ag is mainly the microsomal epoxide hydrolase (mEH), a drug-metabolizing enzyme that plays an important role in the metabolism of some mutagenic and carcinogenic epoxides. Using the purified recombinant human mEH as an antigen, we now found that antibodies against this protein are associated with nearly 82% of hepatitis C virus infections and surprisingly with 46% of patients with hepatitis A. The appearance of AN-Ag/mEH in the incubation period of hepatitis C as previously reported and the antibody responses shown here indicate that this enzyme may be a marker for or even a cause of some of the pathology associated with hepatitis C and A.

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Purification and reconstitution of the bile acid transport system from hepatocyte sinusoidal plasma membranes

Cited by 52 publications

References 30 publications

Cloning and Characterization of a Novel Peptidase from Rat and Human Ileum

Cloning and Characterization of a Novel Peptidase from Rat and Human Ileum

Functional reconstitution of the canalicular bile salt transport system of rat liver.

Autoantibody response to microsomal epoxide hydrolase in hepatitis C and A

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