The Structural Basis for Modular Nonribosomal Peptide Synthesis

 描述(申请人提供)：天然产品是药用活性化合物的丰富来源。一类天然产物是由非核糖体多肽合成酶(NRPS)通过迷人的模块化多结构域结构产生的。在合成过程中，多肽中间体与整合的载体蛋白结合，该载体蛋白以协调的方式输送到邻近的催化域。完全了解NRPS的结构酶学，包括活性部位和结构域界面，将允许通过分析和预测未表征的生物合成簇和将新的化学部分工程到先前已确定的NRPS中来鉴定新产品。为了继续我们对NRPS的结构的询问，我们将确定NRPS多结构域酶的结构，包括具有信息配体的全长模块。我们的研究将涉及执行标准和不寻常多肽合成的多个NRPS系统。我们还将研究增强NRPS腺化结构域的一系列小辅助蛋白的结构基础。最后，我们将研究人类共生细菌的一个新的NRPS系统产生的细胞毒素。这些研究将使用生化、生物物理、结构和化学技术来继续我们的工作，以提供模块NRPS酶的活性位点和结构域的完整结构描述。

 DESCRIPTION (provided by applicant): Natural products are a rich source of pharmaceutically active compounds. One class of natural products are produced by the non-ribosomal peptide synthetases (NRPSs) through a fascinating modular, multidomain architecture. During synthesis, the peptide intermediates are bound to an integrated carrier protein that is delivered in a coordinated manner to neighboring catalytic domains. A complete understanding of the NRPS structural enzymology, including the active sites and the domain interfaces, would allow the identification of novel products both through analysis and prediction of uncharacterized biosynthetic clusters and the engineering of novel chemical moieties into previously identified NRPSs. To continue our structural interrogation of NRPSs, we will determine structures of NRPS multidomain enzymes, including full-length modules, with informative ligands. Our studies will address multiple NRPS systems that perform standard and unusual peptide syntheses. We will additionally examine the structural basis of a family of small auxiliary proteins that enhance NRPS adenylation domains. And finally, we will examine cytotoxin production from a novel NRPS system from a human commensal bacteria. These studies will use biochemical, biophysical, structural, and chemical techniques to continue our work to provide a complete structural description of the active sites and the domain choreography of the modular NRPS enzymes.