Structural analyses of the Group A flavin-dependent monooxygenase PieE reveal a sliding FAD cofactor conformation bridging OUT and IN conformations
文献类型：期刊论文
| 作者 | Manenda, Mahder S.; Picard, Marie-Eve; Zhang, Liping3; Cyr, Normand; Zhu, Xiaojun2; Barma, Julie2; Pascal, John M.; Couture, Manon2; Zhang, Changsheng3; Shi, Rong2 |
| 刊名 | JOURNAL OF BIOLOGICAL CHEMISTRY
 |
| 出版日期 | 2020 |
| 卷号 | 295期号:14页码:4709 |
| 关键词 | flavin adenine dinucleotide (FAD) enzyme catalysis enzyme mechanism small-angle X-ray scattering (SAXS) crystal structure flavin-dependent monooxygenase hydroxylation mobile flavin piericidin sliding conformation |
| ISSN号 | 0021-9258 |
| DOI | 10.1074/jbc.RA119.011212 |
| 英文摘要 | Group A flavin-dependent monooxygenases catalyze the cleavage of the oxygen?oxygen bond of dioxygen, followed by the incorporation of one oxygen atom into the substrate molecule with the aid of NADPH and FAD. These flavoenzymes play an important role in many biological processes, and their most distinct structural feature is the choreographed motions of flavin, which typically adopts two distinct conformations (OUT and IN) to fulfill its function. Notably, these enzymes seem to have evolved a delicate control system to avoid the futile cycle of NADPH oxidation and FAD reduction in the absence of substrate, but the molecular basis of this system remains elusive. Using protein crystallography, size-exclusion chromatography coupled to multi-angle light scattering (SEC-MALS), and small-angle X-ray scattering (SEC-SAXS) and activity assay, we report here a structural and biochemical characterization of PieE, a member of the Group A flavin-dependent monooxygenases involved in the biosynthesis of the antibiotic piericidin A1. This analysis revealed that PieE forms a unique hexamer. Moreover, we found, to the best of our knowledge for the first time, that in addition to the classical OUT and IN conformations, FAD possesses a ?sliding? conformation that exists in between the OUT and IN conformations. This observation sheds light on the underlying mechanism of how the signal of substrate binding is transmitted to the FAD-binding site to efficiently initiate NADPH binding and FAD reduction. Our findings bridge a gap currently missing in the orchestrated order of chemical events catalyzed by this important class of enzymes. |
| 资助机构 | Canada Foundation for InnovationCanada Foundation for Innovation; Natural Sciences and Engineering Research Council of CanadaNatural Sciences and Engineering Research Council of Canada; University of Saskatchewan; Government of Saskatchewan; National Research Council Canada; Canadian Institutes of Health ResearchCanadian Institutes of Health Research (CIHR); Canada Foundation for Innovation AwardCanada Foundation for Innovation [30574] ; Canada Foundation for InnovationCanada Foundation for Innovation; Natural Sciences and Engineering Research Council of CanadaNatural Sciences and Engineering Research Council of Canada; University of Saskatchewan; Government of Saskatchewan; National Research Council Canada; Canadian Institutes of Health ResearchCanadian Institutes of Health Research (CIHR); Canada Foundation for Innovation AwardCanada Foundation for Innovation [30574] |
| 源URL | [http://ir.scsio.ac.cn/handle/344004/18458]  |
| 专题 | 南海海洋研究所_中科院海洋生物资源可持续利用重点实验室 |
| 作者单位 | 1.Cyr, Normand; Pascal, John M.] Univ Montreal, Dept Biochim & Med Mol, Montreal, PQ H3T 1J4, Canada 2.Univ Laval, Dept Biochim Microbiol & Bioinformat, PROTEO, Quebec City, PQ G1V 0A6, Canada 3.Univ Laval, IBIS, Quebec City, PQ G1V 0A6, Canada 4.Chinese Acad Sci, RNAM Ctr Marine Microbiol, South China Sea Inst Oceanol, CAS Key Lab Trop Marine Bioresources & Ecol,Guang, 164 West Xingang Rd, Guangzhou 510301, Peoples R China |
| 推荐引用方式 GB/T 7714 | Manenda, Mahder S.,Picard, Marie-Eve,Zhang, Liping,et al. Structural analyses of the Group A flavin-dependent monooxygenase PieE reveal a sliding FAD cofactor conformation bridging OUT and IN conformations[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2020,295(14):4709, 4722. |
| APA | Manenda, Mahder S..,Picard, Marie-Eve.,Zhang, Liping.,Cyr, Normand.,Zhu, Xiaojun.,...&Shi, Rong.(2020).Structural analyses of the Group A flavin-dependent monooxygenase PieE reveal a sliding FAD cofactor conformation bridging OUT and IN conformations.JOURNAL OF BIOLOGICAL CHEMISTRY,295(14),4709. |
| MLA | Manenda, Mahder S.,et al."Structural analyses of the Group A flavin-dependent monooxygenase PieE reveal a sliding FAD cofactor conformation bridging OUT and IN conformations".JOURNAL OF BIOLOGICAL CHEMISTRY 295.14(2020):4709. |
入库方式： OAI收割
来源：南海海洋研究所
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