Expression, purification, crystallization and preliminary X-ray diffraction studies of the cmcI component of Streptomyces clavuligerus 7alpha-cephem-methoxylase

• Published in: Acta crystallographica. Section D, Biological crystallography. Vol. 60; no. Pt 9; pp. 1618 - 1621
• Main Authors: Lester, Diane R, Oster, Linda M, Svenda, Martin, Andersson, Inger
• Format: Journal Article
• Language: English
• Published: United States 01.09.2004
• Subjects: Crystallization; DNA, Bacterial - chemistry; Electrophoresis, Polyacrylamide Gel; Escherichia coli - metabolism; Methyltransferases - biosynthesis; Methyltransferases - chemistry; Methyltransferases - metabolism; Mixed Function Oxygenases - biosynthesis; Mixed Function Oxygenases - chemistry; Mixed Function Oxygenases - metabolism; Multienzyme Complexes - biosynthesis; Multienzyme Complexes - chemistry; Multienzyme Complexes - metabolism; Protein Conformation; Reverse Transcriptase Polymerase Chain Reaction; Spectrometry, Mass, Electrospray Ionization; Streptomyces - chemistry; Streptomyces - enzymology; X-Ray Diffraction
• ISSN: 0907-4449

Cephamycins are broad-spectrum beta-lactam antibiotics that show resistance to certain forms of beta-lactamases. They differ from cephalosporins by the presence of a methoxyl group at the C-7alpha position. The gene products of cmcI and cmcJ are believed to control 7alpha-methoxylation of cephalosporins through successive steps of hydroxylation and methylation. Here, the expression, purification, crystallization and initial data-collection statistics of the 236-amino-acid protein product of cmcI from Streptomyces clavuligerus is reported. The crystals belong to space group P2(1), with unit-cell parameters a = 93.6, b = 182.6, c = 103.2 A, beta = 91.05 degrees. Diffraction data were collected to 2.5 A.