Takeda Mitsuhiro, Saito Rino, Konno Sho, Nagae Takayuki, Aoyama Hiroshi, Yoshinaga Sosuke, Terasawa Hiroaki, Taguchi Akihiro, Taniguchi Atsuhiko, Hayashi Yoshio, Mishima Masaki
Department of Molecular Biophysics, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan.
Department of Medicinal Chemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences, Tokyo, Japan.
Biomol NMR Assign. 2025 Jun;19(1):17-21. doi: 10.1007/s12104-024-10210-5. Epub 2024 Dec 11.
Natural macrocyclic peptides produced by microorganisms serve as valuable resources for therapeutic compounds, including antibiotics, anticancer agents, and immune suppressive agents. Nonribosomal peptide synthetases (NRPSs) are responsible for the biosynthesis of macrocyclic peptides. NRPSs are large multimodular enzymes, and each module recognizes and incorporates one specific amino acid into the polypeptide product. In the final biosynthetic step, the mature linear peptide precursor is subject to head-to-tail cyclization by the thioesterase (TE) domain in the C-terminal module. Since the TE domains can autonomously catalyze the cyclization of diverse linear peptide substrates, isolated TE domains can be used to produce natural product derivatives. To understand the mechanism of TE domains in NRPSs as a base for therapeutic applications, we investigated the TE domain (residues 6236-6486) of tyrocidine synthetase TycC by NMR. Tyrocidine is a cyclic decapeptide with antibiotic activity, and TycC-TE catalyzes the cyclization of the linear decapeptide precursor. Here, we report the backbone resonance assignments of TycC-TE. The assignments of TycC-TE provide the basis for NMR investigations of the structure and substrate-recognition mode of the TE domain in NRPS.
微生物产生的天然大环肽是治疗性化合物的宝贵资源，包括抗生素、抗癌剂和免疫抑制剂。非核糖体肽合成酶（NRPSs）负责大环肽的生物合成。NRPSs是大型多模块酶，每个模块识别一种特定氨基酸并将其掺入多肽产物中。在最后的生物合成步骤中，成熟的线性肽前体在C端模块中的硫酯酶（TE）结构域作用下进行首尾环化。由于TE结构域可以自主催化多种线性肽底物的环化，因此分离的TE结构域可用于生产天然产物衍生物。为了理解NRPSs中TE结构域的作用机制，作为治疗应用的基础，我们通过核磁共振研究了短杆菌酪肽合成酶TycC的TE结构域（第6236-6486位氨基酸残基）。短杆菌酪肽是一种具有抗生素活性的环十肽，TycC-TE催化线性十肽前体的环化。在此，我们报告了TycC-TE的主链共振归属。TycC-TE的归属为核磁共振研究NRPS中TE结构域的结构和底物识别模式提供了基础。
