Isupov Michail N, Schröder Ewald, Gibson Robert P, Beecher Jean, Donadio Giuliana, Saneei Vahid, Dcunha Stephlina A, McGhie Emma J, Sayer Christopher, Davenport Colin F, Lau Peter C, Hasegawa Yoshie, Iwaki Hiroaki, Kadow Maria, Balke Kathleen, Bornscheuer Uwe T, Bourenkov Gleb, Littlechild Jennifer A
The Henry Wellcome Building for Biocatalysis, Biosciences, College of Life and Environmental Sciences, University of Exeter, Stocker Road, Exeter EX4 4QD, England.
Biotechnology Research Institute, National Research Council Canada, 6100 Royalmount Avenue, Montreal, QC H4P 2R2, Canada.
Acta Crystallogr D Biol Crystallogr. 2015 Nov;71(Pt 11):2344-53. doi: 10.1107/S1399004715017939. Epub 2015 Oct 31.
The three-dimensional structures of the native enzyme and the FMN complex of the overexpressed form of the oxygenating component of the type II Baeyer-Villiger 3,6-diketocamphane monooxygenase have been determined to 1.9 Å resolution. The structure of this dimeric FMN-dependent enzyme, which is encoded on the large CAM plasmid of Pseudomonas putida, has been solved by a combination of multiple anomalous dispersion from a bromine crystal soak and molecular replacement using a bacterial luciferase model. The orientation of the isoalloxazine ring of the FMN cofactor in the active site of this TIM-barrel fold enzyme differs significantly from that previously observed in enzymes of the bacterial luciferase-like superfamily. The Ala77 residue is in a cis conformation and forms a β-bulge at the C-terminus of β-strand 3, which is a feature observed in many proteins of this superfamily.
已确定II型拜耳-维利格3,6-二酮樟脑单加氧酶氧化成分的天然酶和过表达形式的FMN复合物的三维结构，分辨率达到1.9 Å。这种二聚体FMN依赖性酶由恶臭假单胞菌的大型CAM质粒编码，其结构通过溴晶体浸泡的多波长反常散射和使用细菌荧光素酶模型的分子置换相结合的方法得以解析。在这种TIM桶状折叠酶的活性位点中，FMN辅因子的异咯嗪环的取向与先前在细菌荧光素酶样超家族的酶中观察到的取向有显著差异。Ala77残基处于顺式构象，并在β链3的C末端形成一个β凸起，这是在该超家族的许多蛋白质中观察到的一个特征。
