Uptake of organic anions such as bilirubin and sulfobromophthalein (BSP) is a fundamental hepatocyte function. Recently, we cloned a unique mRNA encoding a high affinity organic anion transport protein (OATP). Expression of this protein is sufficient to mediate high affinity Cl-- dependent extraction of 35S-BSP from albumin in Xenopus oocytes and transfected HeLa cells. Preliminary studies reveal that presence of this transport protein is limited to liver and kidney, as determined by Northern blot analysis under high stringency conditions. Morphological studies indicate that OATP is limited to the basolateral (sinusoidal) hepatocyte plasma membrane in normal rat liver. The goal of the proposed studies is to better characterize this transport protein and to understand its mechanism of action. The long-range goal of this work is to understand the mechanism of organic anion transport and its relationship to specific proteins in normal liver and in various acquired and inheritable disorders. To accomplish these goals, three specific aims have been proposed. In the first aim, expression and subcellular localization of OATP will be determined by molecular biological, biochemical, and immunological means. Studies will be focused upon modulation of transport function and mechanisms of regulation of transporter expression in normal, developing and regenerating rat liver.
The second aim will focus studies on the synthesis, post-translational modification, and oligomeric state of OATP The relationship of these modifications to transport function will be elucidated by these studies. The third specific aim will examine the mechanism by which OATP mediates organic anion transport. The substrate binding site will be identified by photoaffinity techniques and its function clarified by studies utilizing site-directed mutagenesis. Activation of transport by Cl- and the role of OH- exchange will be studied in cells stably transfected with OATP.

Agency
National Institute of Health (NIH)
Institute
National Institute of Diabetes and Digestive and Kidney Diseases (NIDDK)
Type
Research Project (R01)
Project #
5R01DK023026-19
Application #
2377730
Study Section
General Medicine A Subcommittee 2 (GMA)
Project Start
1979-03-01
Project End
1999-02-28
Budget Start
1997-03-01
Budget End
1998-02-28
Support Year
19
Fiscal Year
1997
Total Cost
Indirect Cost
Name
Albert Einstein College of Medicine
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
009095365
City
Bronx
State
NY
Country
United States
Zip Code
10461
Bejarano, Eloy; Murray, John W; Wang, Xintao et al. (2018) Defective recruitment of motor proteins to autophagic compartments contributes to autophagic failure in aging. Aging Cell :e12777
Murray, John W; Yin, David; Wolkoff, Allan W (2017) Reduction of organelle motility by removal of potassium and other solutes. PLoS One 12:e0184898
Roy-Chowdhury, Jayanta; Roy-Chowdhury, Namita; Listowsky, Irving et al. (2017) Drug- and Drug Abuse-Associated Hyperbilirubinemia: Experience With Atazanavir. Clin Pharmacol Drug Dev 6:140-146
Wang, Xintao; Wang, Pijun; Wang, Wenjun et al. (2016) The Na(+)-Taurocholate Cotransporting Polypeptide Traffics with the Epidermal Growth Factor Receptor. Traffic 17:230-44
Murray, John W; Han, Dennis; Wolkoff, Allan W (2014) Hepatocytes maintain greater fluorescent bile acid accumulation and greater sensitivity to drug-induced cell death in three-dimensional matrix culture. Physiol Rep 2:
Wang, Wen-Jun; Murray, John W; Wolkoff, Allan W (2014) Oatp1a1 requires PDZK1 to traffic to the plasma membrane by selective recruitment of microtubule-based motor proteins. Drug Metab Dispos 42:62-9
Yuan, Fei; Snapp, Erik L; Novikoff, Phyllis M et al. (2014) Human liver cell trafficking mutants: characterization and whole exome sequencing. PLoS One 9:e87043
Wolkoff, Allan W (2014) Organic anion uptake by hepatocytes. Compr Physiol 4:1715-35
Mukhopadhyay, Aparna; Quiroz, Jose A; Wolkoff, Allan W (2014) Rab1a regulates sorting of early endocytic vesicles. Am J Physiol Gastrointest Liver Physiol 306:G412-24
Choi, Jo H; Murray, John W; Wolkoff, Allan W (2011) PDZK1 binding and serine phosphorylation regulate subcellular trafficking of organic anion transport protein 1a1. Am J Physiol Gastrointest Liver Physiol 300:G384-93

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