Crystallization and preliminary X-ray diffraction analysis of three recombinant mutants of Vaccinia virus uracil DNA glycosylase.
Publication Type:Journal Article
Source:Acta Crystallogr Sect F Struct Biol Cryst Commun, Volume 69, Issue Pt 3, p.295-301 (2013)
Keywords:Amino Acid Sequence, Amino Acids, Crystallization, Crystallography, X-Ray, Escherichia coli, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Protein Structure, Secondary, Protein Structure, Tertiary, Recombinant Proteins, Uracil-DNA Glycosidase, Vaccinia virus, Viral Proteins
<p>Amino-acid residues located at a highly flexible area in the uracil DNA glycosylase of Vaccinia virus were mutated. In the crystal structure of wild-type D4 these residues lie at the dimer interface. Specifically, three mutants were generated: (i) residue Arg167 was replaced with an alanine (R167AD4), (ii) residues Glu171, Ser172 and Pro173 were substituted with three glycine residues (3GD4) and (iii) residues Glu171 and Ser172 were deleted (Δ171-172D4). Mutant proteins were expressed, purified and crystallized in order to investigate the effects of these mutations on the structure of the protein.</p>