Time-resolved crystallographic and spectroscopic studies of the mononuclear non-haem iron oxygenases, IPNS and VioC

单核非血红素铁加氧酶、IPNS 和 VioC 的时间分辨晶体学和光谱研究

• 批准号: 2107984
• 金额: --
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2107984
• 关键词: Time; resolved; crystallographic; spectroscopic; studies

Research within the field of structural biology aims to provide full understanding of the structure andfunction of biological macromolecules, information that ultimately facilitates rational drug design andprotein engineering. Application of structural biology to the class of mononuclear non-haem ironoxygenases has potential to help combat the growing problem of antibiotic resistance. These enzymesare responsible for the incorporation of molecular oxygen (O2) into a wide variety of substrates. Dueto the intrinsic properties of dioxygen, reactions are difficult but biologically rewarding, exemplifiedby the number of enzymes within the family and the range of functions they facilitate. Two structurallysimilar enzymes from this class are isopenicillin N synthase (IPNS) and viomycin biosynthetic proteinC. The former catalyses the of bicyclic ring closure forming the B-lactam core of penicillin andcephalosporin antibiotics. Viomycin biosynthetic protein C (VioC) catalyses hydroxylation of L-arginine(L-arg), which is subsequently cyclised forming the non-standard amino acid L-capreomycidine (L-cam)and incorporated into the peptide precursor of viomycin. While structurally similar, IPNS and VioC aremechanistically distinct. VioC catalysis requires the presence of a cofactor (a-ketoglutarate) while theIPNS reaction is cofactor-independent. Due to the importance of this class of enzymes, both proteinshave been subject to extensive research. Nevertheless, questions of their precise reactionmechanisms remain unanswered. The aim of this project is to fully elucidate the mechanisms of bothenzymes. The project will involve the use of time-resolved crystallographic studies made possible byX-ray free electron laser facilities. Due to structural similarity within the oxygenase family, theinformation collected from these enzymes will provide understanding of a wide range of biologicalrelevant oxygenase proteins.

结构生物学领域的研究旨在提供对生物大分子的结构和功能的充分了解，这些信息最终有助于合理的药物设计和蛋白质工程。将结构生物学应用于单核非血红素铁加氧酶类，有可能帮助解决日益严重的抗生素耐药性问题。这些酶负责将分子氧(O2)结合到各种底物中。由于氧气的固有性质，反应是困难的，但在生物学上是有益的，例如家族中的酶的数量和它们促进的一系列功能。两种结构相似的酶是异青霉素N合成酶(IPNS)和万古霉素生物合成蛋白C。前者催化双环闭合形成青霉素和头孢菌素类抗生素的B-内酰胺核心。紫霉素生物合成蛋白C(VioC)催化L-精氨酸(L-Arg)的羟基化，进而循环形成非标准氨基酸L-卷曲霉菌素(L-CaM)，并被引入到维霉素肽的前体中。虽然IPNS和VioC在结构上相似，但在机制上是不同的。VioC催化需要辅因子(α-酮戊二酸)的存在，而IPNS反应不依赖辅因子。由于这类酶的重要性，这两种蛋白质都受到了广泛的研究。然而，关于它们的确切反应机制的问题仍然没有得到回答。本项目的目的是全面阐明这两种酶的作用机制。该项目将使用时间分辨的结晶学研究，使X射线自由电子激光设备成为可能。由于加氧酶家族的结构相似，从这些酶中收集的信息将提供对广泛的生物相关加氧酶蛋白的理解。