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Production and crystallization of a selenomethionyl variant of UmuD′, an Echerichia coli SOS response protein
Author(s) -
Peat Thomas S.,
Frank Ekaterina G.,
Woodgate Roger,
Hendrickson Wayne A.
Publication year - 1996
Publication title -
proteins: structure, function, and bioinformatics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.699
H-Index - 191
eISSN - 1097-0134
pISSN - 0887-3585
DOI - 10.1002/(sici)1097-0134(199608)25:4<506::aid-prot10>3.0.co;2-l
Subject(s) - methionine , crystallization , mutant , amino acid , valine , escherichia coli , protein crystallization , threonine , mutagenesis , chemistry , crystallography , residue (chemistry) , selenium , tyrosine , cysteine , biochemistry , stereochemistry , organic chemistry , serine , phosphorylation , enzyme , gene
Crystals of both native and mutant Escherichia coli UmuD′ protein were obtained using the hanging drop method. Soaking the native crystals in solutions of heavy metal ions failed to produce good isomorphous derivatives, and selenomethionine substituted wild‐type protein did not crystallize under conditions that gave native crystals. Site‐directed mutagenesis was used to change the penultimate residue, a methionine amino acid, to either a valine or a threonine amino acid. Crystals were subsequently obtained from these mutant proteins with and without selenomethionine Incorporation. Crystals of the native, the mutant, and the selenomethionine Incorporated protein were all similar, crystallizing in the P4 1 2 1 2 space group. © 1996 Wiley‐Liss, Inc.