The Structural Basis for Modular Nonribosomal Peptide Synthesis

 描述（由申请人提供）：天然产品是药物活性化合物的丰富来源。一类天然产物是由非核糖体肽合成酶（NRPS）通过迷人的模块化多结构域结构产生的。在合成过程中，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递送至相邻的催化结构域。NRPS结构酶学，包括活性位点和结构域接口的完整理解，将允许通过分析和预测的未表征的生物合成簇和工程的新的化学部分到以前确定的NRPS的新产品的识别。为了继续我们对NRPS的结构询问，我们将确定NRPS多结构域酶的结构，包括全长模块和信息配体。我们的研究将解决多个NRPS系统，执行标准和不寻常的肽合成。我们还将研究增强NRPS腺苷酸化结构域的小辅助蛋白家族的结构基础。最后，我们将研究从一个新的NRPS系统从人类肠道细菌的细胞毒素的生产。这些研究将使用生物化学，生物物理，结构和化学技术，继续我们的工作，提供一个完整的结构描述的活性位点和模块化NRPS酶的结构域编排。

 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.