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Crystallization and preliminary X‐ray investigation of a recombinant form of rat catechol O ‐methyltransferase
Author(s) -
Vidgren Jukka,
Tilgmann Carola,
Lundström Kenneth,
Liljas Anders
Publication year - 1991
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/prot.340110309
Subject(s) - recombinant dna , polyethylene glycol , crystallization , catechol , chemistry , enzyme , methyltransferase , molecule , o methyltransferase , complementary dna , microbiology and biotechnology , biochemistry , biology , dna , organic chemistry , methylation , gene
Rat catechol O ‐methyltransferase cDNA was introduced into an E. coli expression vector pKEX14, which utilizes the inducible T7 promoter. Active and soluble recombinant catechol O ‐methyltransferase was produced in bacteria and purified to electrophoretic homogeneity by chromatographic procedures. The purified enzyme has been crystallized by the method of vapor diffusion using polyethylene glycol as precipitant. The space group is P 3 1 21 or P 3 2 21 with a = b = 51.3 Å and c = 168.5 Å and one molecule in the asymmetric unit. The crystals diffract beyond 3.2 Å and are suitable for three‐dimensional X‐ray structure determination.