Your browser doesn't support javascript.
loading
Functional characterization of human equilibrative nucleoside transporter 1
Protein & Cell ; (12): 284-295, 2017.
Article in English | 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.
Subject(s)

Full text: Available Index: WPRIM (Western Pacific) Main subject: Structure-Activity Relationship / Recombinant Proteins / Adenine Nucleotides / Chemistry / Equilibrative Nucleoside Transporter 1 / HEK293 Cells / Protein Domains / Genetics / Metabolism Limits: Humans Language: English Journal: Protein & Cell Year: 2017 Type: Article

Similar

MEDLINE

...
LILACS

LIS

Full text: Available Index: WPRIM (Western Pacific) Main subject: Structure-Activity Relationship / Recombinant Proteins / Adenine Nucleotides / Chemistry / Equilibrative Nucleoside Transporter 1 / HEK293 Cells / Protein Domains / Genetics / Metabolism Limits: Humans Language: English Journal: Protein & Cell Year: 2017 Type: Article