AUTHOR=Hao Zhenyu , Thomsen Maren , Postis Vincent L. G. , Lesiuk Amelia , Sharples David , Wang Yingying , Bartlam Mark , Goldman Adrian 

TITLE=A Novel and Fast Purification Method for Nucleoside Transporters

JOURNAL=Frontiers in Molecular Biosciences

VOLUME=3

YEAR=2016

URL=https://www.frontiersin.org/journals/molecular-biosciences/articles/10.3389/fmolb.2016.00023

DOI=10.3389/fmolb.2016.00023

ISSN=2296-889X

ABSTRACT=<p>Nucleoside transporters (NTs) play critical biological roles in humans, and to understand the molecular mechanism of nucleoside transport requires high-resolution structural information. However, the main bottleneck for structural analysis of NTs is the production of pure, stable, and high quality native protein for crystallization trials. Here we report a novel membrane protein expression and purification strategy, including construction of a high-yield membrane protein expression vector, and a new and fast purification protocol for NTs. The advantages of this strategy are the improved time efficiency, leading to high quality, active, stable membrane proteins, and the efficient use of reagents and consumables. Our strategy might serve as a useful point of reference for investigating NTs and other membrane proteins by clarifying the technical points of vector construction and improvements of membrane protein expression and purification.</p>