Untersuchung der Wechselwirkung von PCP Domänen mit Interaktionspartnern in Nicht-ribosomalen Peptidsynthetasen

PCP 结构域与非核糖体肽合成酶中相互作用伙伴的相互作用的研究

• 批准号: 171029658
• 负责人: Professor Dr. Volker Dötsch
• 金额: --
• 依托单位: Institut für Biophysikalische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2014-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/171029658?language=en
• 关键词: Untersuchung; der; Wechselwirkung; von; PCP

Non-ribosomal peptide synthetases (NRPS) constitute a class of large multidomain enzymes found in the cytoplasm of bacteria and fungi that produce a large set of peptides with biological activities ranging from iron sequestration to chemical defense against other microbes. Many of these peptides are used as drugs with antibiotic, anticancer or immunosuppressive activity, making NRPS systems of high medical importance. The modular structure of these NRPS systems has fueled attempts to create new, potentially medically important peptides by combining modules from different NRPS systems into new assembly lines. However, many design experiments have shown that even small changes in the assembly lines result in significant reductions in yield, suggesting that the individual modules are not mere beads on a string but that specific domaindomain interactions play important roles. In previous studies we investigated the interaction of peptide carrier proteins (PCPs) with thioesterases and other intrans interacting enzymes. In this grant application we describe detailed studies of the domain-domain interaction of PCPs with the major components of NRPS systems, the A- and the C-domains that are responsible for amino acids type selection and for performing the chemical condensation step of the reaction. In particular we will focus on the role of peptides that are covalently attached to the PCP and that modulate the interaction with C-domains.

非核糖体肽合成酶（NRPS）是一类存在于细菌和真菌细胞质中的大的多结构域酶，其产生大量具有从铁螯合到针对其他微生物的化学防御的生物活性的肽。这些肽中的许多被用作具有抗生素、抗癌或免疫抑制活性的药物，使得NRPS系统具有高度的医学重要性。这些NRPS系统的模块化结构推动了通过将来自不同NRPS系统的模块组合到新的装配线中来创建新的、潜在的医学重要肽的尝试。然而，许多设计实验表明，即使是在装配线上的小变化也会导致产量的显着降低，这表明单个模块不仅仅是一串珠子，而是特定的domaindomain相互作用起着重要的作用。在以前的研究中，我们研究了肽载体蛋白（PCP）与硫酯酶和其他反式相互作用酶的相互作用。在本授权申请中，我们描述了PCP与NRPS系统的主要组分，负责氨基酸类型选择和进行反应的化学缩合步骤的A-和C-结构域的结构域-结构域相互作用的详细研究。特别是，我们将集中在共价连接到PCP和调节与C-结构域的相互作用的肽的作用。

项目成果

Assembling a Correctly Folded and Functional Heptahelical Membrane Protein by Protein Trans-splicing*

通过蛋白质反式拼接组装正确折叠且具有功能的七螺旋膜蛋白质*

• DOI: 10.1074/jbc.m115.681205
• 发表时间: 2015
• 期刊: The Journal of Biological Chemistry
• 作者: Mehler M;Eckert CE;Busche A;Kulhei J;Michaelis J;Becker-Baldus J;Wachtveitl J;Dötsch V;Glaubitz C
• 通讯作者: Glaubitz C

SPLICEFINDER – A Fast and Easy Screening Method for Active Protein Trans-Splicing Positions

SPLICEFINDER – 一种快速简便的活性蛋白反式剪接位置筛选方法

• DOI: 10.1371/journal.pone.0072925
• 发表时间: 2013
• 期刊: PLoS ONE
• 影响因子: 3.7
• 作者: Zettler J;Eppmann S;Busche A;Dikovskaya D;Dötsch V;Mootz HD;Sonntag T
• 通讯作者: Sonntag T