Purification, crystallization and preliminary X-ray diffraction analysis of the human mismatch repair protein MutS?.

ABSTRACT: MutS? is a eukaryotic mismatch repair protein that preferentially targets extrahelical unpaired nucleotides and shares partial functional redundancy with MutS? (MSH2-MSH6). Although mismatch recognition by MutS? has been shown to involve a conserved Phe-X-Glu motif, little is known about the lesion-binding mechanism of MutS?. Combined MSH3/MSH6 deficiency triggers a strong predisposition to cancer in mice and defects in msh2 and msh6 account for roughly half of hereditary nonpolyposis colorectal cancer mutations. These three MutS homologs are also believed to play a role in trinucleotide repeat instability, which is a hallmark of many neurodegenerative disorders. The baculovirus overexpression and purification of recombinant human MutS? and three truncation mutants are presented here. Binding assays with heteroduplex DNA were carried out for biochemical characterization. Crystallization and preliminary X-ray diffraction analysis of the protein bound to a heteroduplex DNA substrate are also reported.

SUBMITTER: Tseng Q 

PROVIDER: S-EPMC3151135 | biostudies-literature | 2011 Aug

REPOSITORIES: biostudies-literature