6HNL
Selenomethionine derivative of IdmH 96-104 loop truncation variant
6HNL の概要
エントリーDOI | 10.2210/pdb6hnl/pdb |
関連するPDBエントリー | 6HNM 6HNN |
分子名称 | Putative polyketide cyclase IdmH (2 entities in total) |
機能のキーワード | polyketide synthesis, putative cyclase, biosynthetic protein |
由来する生物種 | Streptomyces antibioticus |
タンパク質・核酸の鎖数 | 2 |
化学式量合計 | 30934.86 |
構造登録者 | Drulyte, I.,Obajdin, J.,Trinh, C.,Hemsworth, G.R.,Berry, A. (登録日: 2018-09-16, 公開日: 2019-11-06, 最終更新日: 2024-11-06) |
主引用文献 | Drulyte, I.,Obajdin, J.,Trinh, C.H.,Kalverda, A.P.,van der Kamp, M.W.,Hemsworth, G.R.,Berry, A. Crystal structure of the putative cyclase IdmH from the indanomycin nonribosomal peptide synthase/polyketide synthase. Iucrj, 6:1120-1133, 2019 Cited by PubMed Abstract: Indanomycin is biosynthesized by a hybrid nonribosomal peptide synthase/polyketide synthase (NRPS/PKS) followed by a number of 'tailoring' steps to form the two ring systems that are present in the mature product. It had previously been hypothesized that the indane ring of indanomycin was formed by the action of IdmH using a Diels-Alder reaction. Here, the crystal structure of a selenomethionine-labelled truncated form of IdmH (IdmH-Δ99-107) was solved using single-wavelength anomalous dispersion (SAD) phasing. This truncated variant allows consistent and easy crystallization, but importantly the structure was used as a search model in molecular replacement, allowing the full-length IdmH structure to be determined to 2.7 Å resolution. IdmH is a homodimer, with the individual protomers consisting of an α+β barrel. Each protomer contains a deep hydrophobic pocket which is proposed to constitute the active site of the enzyme. To investigate the reaction catalysed by IdmH, 88% of the backbone NMR resonances were assigned, and using chemical shift perturbation of [N]-labelled IdmH it was demonstrated that indanomycin binds in the active-site pocket. Finally, combined quantum mechanical/molecular mechanical (QM/MM) modelling of the IdmH reaction shows that the active site of the enzyme provides an appropriate environment to promote indane-ring formation, supporting the assignment of IdmH as the key Diels-Alderase catalysing the final step in the biosynthesis of indanomycin through a similar mechanism to other recently characterized Diels-Alderases involved in polyketide-tailoring reactions. An animated Interactive 3D Complement (I3DC) is available in Proteopedia at https://proteopedia.org/w/Journal:IUCrJ:S2052252519012399. PubMed: 31709067DOI: 10.1107/S2052252519012399 主引用文献が同じPDBエントリー |
実験手法 | X-RAY DIFFRACTION (2.2 Å) |
構造検証レポート
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