Design, synthesis, and evaluation of 5'-S-aminoethyl-N(6)- azidobenzyl-5'-thioadenosine biotin conjugate: a bifunctional photoaffinity probe for the es nucleoside transporter

Bioconjug Chem. 2004 May-Jun;15(3):536-40. doi: 10.1021/bc034165j.

Abstract

A bifunctional biotinylated photoaffinity label for the nitrobenzylmercaptopurine riboside (NBMPR)-sensitive (es) nucleoside transporter (ENT1) has been synthesized and evaluated. This new probe,5'-S-aminoethyladenosine-N(6)-azidobenzyl-5'-thioadenosine biotin conjugate (SAEATA-14-biotin), exhibited high-affinity binding to the es transporter in K562 cells as determined by flow cytometry, with a K(i) of 2.69 nM in competition against 5-(SAENTA)-x8-fluorescein. It also exhibited covalent linking to the es transporter in BeWo cell membranes upon UV irradiation. This new bifunctional probe is a potential tool for determining the amino acid residues involved in ligand binding at the NBMPR-binding site of the ENT1 nucleoside transporter, as well as for the purification of the transporter.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adenosine / analogs & derivatives*
  • Adenosine / chemical synthesis
  • Adenosine / chemistry*
  • Biotin / chemical synthesis
  • Biotin / chemistry*
  • Drug Design
  • Ligands
  • Molecular Structure
  • Nucleoside Transport Proteins / chemistry
  • Nucleoside Transport Proteins / drug effects
  • Nucleoside Transport Proteins / metabolism*
  • Photoaffinity Labels / chemistry*
  • Photoaffinity Labels / metabolism*
  • Photoaffinity Labels / pharmacokinetics
  • Thionucleosides / chemical synthesis
  • Thionucleosides / chemistry*

Substances

  • Ligands
  • Nucleoside Transport Proteins
  • Photoaffinity Labels
  • Thionucleosides
  • 5'-S-(2-aminoethyl)-N(6)-(4-nitrobenzyl)-5'-thioadenosine
  • Biotin
  • Adenosine