An improved procedure for the preparation of X-ray diffraction-quality crystals of cytochrome p450cam.
A procedure for the crystallization of recombinant cytochrome P450cam has been developed which avoids the difficulties inherent in the glass-capillary free-interface diffusion method reported previously. The surface mutation, Cys334-->Ala (C334A), originally designed to prevent dimer formation and thus improve routine handling of the enzyme, facilitates crystallization by the hanging-drop vapour-diffusion technique. Crystals of (C334A)P450cam grow within 48 h and diffract to beyond 1.2 A at 100 K in-house on a Siemens multiwire area detector. Data have been collected from the camphor-bound form to 1.35 A.
Department of Chemistry, Inorganic Chemistry Laboratory, University of Oxford, Oxford OX1 3QR, England. darren.nickerson@icl.ox.ac.uk
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D000409:Alanine; D019943:Amino Acid Substitution; D019475:Camphor 5-Monooxygenase / Q000737:chemistry* / Q000235:genetics; D003545:Cysteine; D017354:Point Mutation; D011994:Recombinant Proteins / Q000737:chemistry; D014961:X-Ray Diffraction
D011994:Recombinant Proteins; D019475:Camphor 5-Monooxygenase; D003545:Cysteine; D000409:Alanine