Schaub Andrew J, Moreno Gabriel O, Zhao Shiji, Truong Hau V, Luo Ray, Tsai Shiou-Chuan
Department of Chemistry, University of California, Irvine, CA, United States.
Department of Molecular Biology and Biochemistry, University of California, Irvine, CA, United States.
Methods Enzymol. 2019;622:375-409. doi: 10.1016/bs.mie.2019.03.001. Epub 2019 Apr 22.
Various computational methodologies can be applied to enzymological studies on enzymes in the fatty acid, polyketide, and non-ribosomal peptide biosynthetic pathways. These multi-domain complexes are called fatty acid synthases, polyketide synthases, and non-ribosomal peptide synthetases. These mega-synthases biosynthesize chemically diverse and complex bioactive molecules, with the intermediates being chauffeured between catalytic partners via a carrier protein. Recent efforts have been made to engineer these systems to expand their product diversity. A major stumbling block is our poor understanding of the transient protein-protein and protein-substrate interactions between the carrier protein and its many catalytic partner domains and product intermediates. The innate reactivity of pathway intermediates in two major classes of polyketide synthases has frustrated our mechanistic understanding of these interactions during the biosynthesis of these natural products, ultimately impeding the engineering of these systems for the generation of engineered natural products. Computational techniques described in this chapter can aid data interpretation or used to generate testable models of these experimentally intractable transient interactions, thereby providing insight into key interactions that are difficult to capture otherwise, with the potential to expand the diversity in these systems.
各种计算方法可应用于脂肪酸、聚酮化合物和非核糖体肽生物合成途径中酶的酶学研究。这些多结构域复合物被称为脂肪酸合酶、聚酮化合物合酶和非核糖体肽合成酶。这些巨型合酶生物合成化学性质多样且复杂的生物活性分子,其中间体通过载体蛋白在催化伙伴之间传递。最近人们努力对这些系统进行工程改造以扩大其产物多样性。一个主要障碍是我们对载体蛋白与其众多催化伙伴结构域和产物中间体之间的瞬时蛋白质 - 蛋白质和蛋白质 - 底物相互作用了解不足。两类主要聚酮化合物合酶中途径中间体的固有反应性阻碍了我们对这些天然产物生物合成过程中这些相互作用的机制理解,最终妨碍了对这些系统进行工程改造以生成工程化天然产物。本章中描述的计算技术可帮助解释数据或用于生成这些难以通过实验处理的瞬时相互作用的可测试模型,从而深入了解难以通过其他方式捕捉的关键相互作用,有可能扩大这些系统中的多样性。