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) 是细菌和真菌细胞质中发现的一类大型多域酶，可产生大量具有从铁螯合到针对其他微生物的化学防御等生物活性的肽。许多这些肽被用作具有抗生素、抗癌或免疫抑制活性的药物，使得 NRPS 系统具有很高的医学重要性。这些 NRPS 系统的模块化结构推动了通过将不同 NRPS 系统的模块组合到新的装配线中来创造新的、具有潜在医学重要性的肽的尝试。然而，许多设计实验表明，即使装配线中的微小变化也会导致产量显着降低，这表明各个模块不仅仅是串上的珠子，而且特定的域域相互作用发挥着重要作用。在之前的研究中，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。在这项拨款申请中，我们描述了 PCP 与 NRPS 系统主要成分（负责氨基酸类型选择和执行反应化学缩合步骤的 A 和 C 结构域）的结构域相互作用的详细研究。我们将特别关注共价连接到 PCP 并调节与 C 结构域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