title: Purification, crystallization and preliminary X-ray diffraction analysis of the human mismatch repair protein MutSβ.
abstract: MutSβ is a eukaryotic mismatch repair protein that preferentially targets extrahelical unpaired nucleotides and shares partial functional redundancy with MutSα (MSH2-MSH6). Although mismatch recognition by MutSα has been shown to involve a conserved Phe-X-Glu motif, little is known about the lesion-binding mechanism of MutSβ. 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 MutSβ 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.
authors.affiliation: Department of Biochemistry, Duke University Medical Center, Durham, NC 27710, USA.
authors.forename: Quincy
authors.initials: Q
authors.lastname: Tseng
authors.forename: Jillian
authors.initials: J
authors.lastname: Orans
authors.forename: Michael A
authors.initials: MA
authors.lastname: Hast
authors.forename: Ravi R
authors.initials: RR
authors.lastname: Iyer
authors.forename: Anita
authors.initials: A
authors.lastname: Changela
authors.forename: Paul L
authors.initials: PL
authors.lastname: Modrich
authors.forename: Lorena S
authors.initials: LS
authors.lastname: Beese
mesh_terms: D003460:Crystallization; D018360:Crystallography, X-Ray; D006801:Humans; D051718:MutS Homolog 2 Protein / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification; D009154:Mutation; D011485:Protein Binding
chemical_list: D051718:MutS Homolog 2 Protein