Date of Award

2011

Degree Type

Thesis

Degree Name

Master of Science

Program

Pharmacology and Toxicology

Supervisor

Dr. James R. Hammond

Abstract

Site-directed mutagenesis was used to target residues within transmembrane domains 1 and 2 of the human equilibrative nucleoside transporter 1 (hENTl), in order to assess their contributions to transporter functionality. Through affinity changes in [ H]NBMPR binding, competitive inhibition profiles with ENT1 inhibitors and [ H]2- chloroadenosine uptake, these residues were found to make minor, but statistically significant, contributions to the inhibitor and/or substrate binding site of hENTl.

An initial study was also undertaken to probe the potential oligomerization of hENTl. Typically considered to be a monomer to this date, under the native conditions of Blue Native Electrophoresis, hENTl was found to possess a molecular mass of 160 kDa. This is approximately three times the size of the known 55kDa hENTl monomer. Targeted site-directed mutagenesis of GxxxG motifs within the transporter, known to play a role in oligomerization, did not appear to affect the higher molecular mass hENTl complex observed

Share

COinS
 
 

To view the content in your browser, please download Adobe Reader or, alternately,
you may Download the file to your hard drive.

NOTE: The latest versions of Adobe Reader do not support viewing PDF files within Firefox on Mac OS and if you are using a modern (Intel) Mac, there is no official plugin for viewing PDF files within the browser window.