The Blood-Brain Barrier Thyroxine Transporter Organic Anion-Transporting Polypeptide 1c1 Displays Atypical Transport Kinetics
| Autor: | Jon N. Rumbley, David R. Salo, Grant W. Anderson, Kevin J. Viken, Daniel E. Westholm |
|---|---|
| Rok vydání: | 2009 |
| Předmět: |
medicine.medical_specialty
Organic Cation Transport Proteins Stereochemistry Kinetics Plasma protein binding Blood–brain barrier Cell Line Endocrinology Internal medicine medicine Animals Humans Binding site Estradiol biology Chemistry Substrate (chemistry) Biological Transport Transporter Rats Organic anion-transporting polypeptide Thyroxine medicine.anatomical_structure Blood-Brain Barrier biology.protein Protein Binding Organic anion |
| Zdroj: | Endocrinology. 150:5153-5162 |
| ISSN: | 1945-7170 0013-7227 |
| DOI: | 10.1210/en.2009-0769 |
| Popis: | Organic anion-transporting polypeptide (Oatp) 1c1 is a high-affinity T(4) transporter expressed in brain barrier cells. Oatp1c1 transports a variety of additional ligands including the conjugated sterol estradiol 17beta-glucuronide (E(2)17betaG). Intriguingly, published data suggest that E(2)17betaG inhibition of Oatp1c1-mediated T(4) transport exhibits characteristics suggestive of atypical transport kinetics. To determine whether Oatp1c1 exhibits atypical transport kinetics, we first performed detailed T(4) and E(2)17betaG uptake assays using Oatp1c1 stably transfected HEK293 cells and a wide range of T(4) and E(2)17betaG concentrations (100 pm to 300 nm and 27 nm to 200 mum, respectively). Eadie-Hofstee plots derived from these detailed T(4) and E(2)17betaG uptake experiments display a biphasic profile consistent with atypical transport kinetics. These data along with T(4) and E(2)17betaG cis-inhibition dose-response measurements revealed shared high- and low-affinity Oatp1c1 binding sites for T(4) and E(2)17betaG. T(4) and E(2)17betaG recognized these Oatp1c1 binding sites with opposite preferences. In addition, sterols glucuronidated in the 17 or 21 position, exhibited preferential substrate-dependent inhibition of Oatp1c1 transport, inhibiting Oatp1c1-mediated E(2)17betaG transport more strongly than T(4) transport. Together these data reveal that Oatp1c1-dependent substrate transport is a complex process involving substrate interaction with multiple binding sites and competition for binding with a variety of other substrates. A thorough understanding of atypical Oatp1c1 transport processes and substrate-dependent inhibition will allow better prediction of endo- and xenobiotic interactions with the Oatp transporter. |
| Databáze: | OpenAIRE |
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