Functional characterization of human equilibrative nucleoside transporter 1
Protein & Cell
;
(12): 284-295, 2017.
Artículo
en Inglés
| WPRIM
| ID: wpr-757343
ABSTRACT
Equilibrative nucleoside transporters (ENTs), which facilitate cross-membrane transport of nucleosides and nucleoside-derived drugs, play an important role in the salvage pathways of nucleotide synthesis, cancer chemotherapy, and treatment for virus infections. Functional characterization of ENTs at the molecular level remains technically challenging and hence scant. In this study, we report successful purification and biochemical characterization of human equilibrative nucleoside transporter 1 (hENT1) in vitro. The HEK293F-derived, recombinant hENT1 is homogenous and functionally active in proteoliposome-based counter flow assays. hENT1 transports the substrate adenosine with a K of 215 ± 34 µmol/L and a V of 578 ± 23.4 nmol mg min. Adenosine uptake by hENT1 is competitively inhibited by nitrobenzylmercaptopurine ribonucleoside (NBMPR), nucleosides, deoxynucleosides, and nucleoside-derived anti-cancer and anti-viral drugs. Binding of hENT1 to adenosine, deoxyadenosine, and adenine by isothermal titration calorimetry is in general agreement with results of the competitive inhibition assays. These results validate hENT1 as a bona fide target for potential drug target and serve as a useful basis for future biophysical and structural studies.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Relación Estructura-Actividad
/
Proteínas Recombinantes
/
Nucleótidos de Adenina
/
Química
/
Tranportador Equilibrativo 1 de Nucleósido
/
Células HEK293
/
Dominios Proteicos
/
Genética
/
Metabolismo
Límite:
Humanos
Idioma:
Inglés
Revista:
Protein & Cell
Año:
2017
Tipo del documento:
Artículo
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