Purification of a Multidrug Resistance Transporter for Crystallization Studies.

ABSTRACT: Crystallization of integral membrane proteins is a challenging field and much effort has been invested in optimizing the overexpression and purification steps needed to obtain milligram amounts of pure, stable, monodisperse protein sample for crystallography studies. Our current work involves the structural and functional characterization of the Escherichia coli multidrug resistance transporter MdtM, a member of the major facilitator superfamily (MFS). Here we present a protocol for isolation of MdtM to increase yields of recombinant protein to the milligram quantities necessary for pursuit of structural studies using X-ray crystallography. Purification of MdtM was enhanced by introduction of an elongated His-tag, followed by identification and subsequent removal of chaperonin contaminatio

SUBMITTER: Alegre KO 

PROVIDER: S-EPMC4790320 | biostudies-literature | 2015 Mar

REPOSITORIES: biostudies-literature