Preliminary crystallographic analysis of an enzyme involved in erythromycin biosynthesis: cytochrome P450eryF.
Cytochrome P450eryF was overexpressed in Escherichia coli and purified in high yield. Crystals of the protein in the presence of the substrate, 6-deoxyerythronolide B, have been obtained by the hanging drop vapor diffusion method, using polyethylene glycol 4000 as a precipitant. The crystals belong to the orthorhombic space group P2(1)2(1)2(1) with unit cell dimensions of a = 54.16 A, b = 79.67 A, and c = 99.48 A and one molecule per asymmetric unit. A complete native data set has been collected to a resolution of 2.1 A, and anomalous dispersion difference Patterson maps have revealed the location of the single heme iron atom.
Department of Molecular Biology and Biochemistry, University of California, Irvine 92717.
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Cupp-Vickery
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Li
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TL
Poulos
D000595:Amino Acid Sequence; D001426:Bacterial Proteins; D018360:Crystallography, X-Ray; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D004917:Erythromycin / Q000031:analogs & derivatives / Q000096:biosynthesis / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D006899:Mixed Function Oxygenases / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D011994:Recombinant Proteins / Q000737:chemistry; D016930:Saccharopolyspora / Q000201:enzymology* / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D001426:Bacterial Proteins; D011994:Recombinant Proteins; C034277:6-deoxyerythronolide B; C038315:erythronolide B; D004917:Erythromycin; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; C054444:eryF protein, Saccharopolyspora erythraea