Structure determination of the membrane bound protease LspA by NMR spectroscopy

核磁共振波谱法测定膜结合蛋白酶 LspA 的结构

• 批准号: 261919446
• 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: 2014
• 资助国家: 德国
• 起止时间: 2013-12-31 至 2015-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/261919446?language=en
• 关键词: Structure; determination; membrane; bound; protease

Lipoprotein signal peptidase II (LspA) is a bacterial protease located in the bacterial membrane. LspA cleaves the signal peptide from prolipoproteins after they are exported across the inner membrane. LspA belongs to an unusual class of aspartic acid proteases with no homologous proteins present in archaea or eukaryotes which makes it an excellent target for the development of new antibiotics. The protein contains 164 amino acids with four predicted alpha-helices. We want to determine the structure of LspA by liquid state NMR spectroscopy and investigate how the cyclic peptide inhibitor globomycin interacts with the protein. We have established expression and labelling protocols using our cell-free expression system and have measured 2D and 3D NMR spectra of LspA in nanodiscs that show very good chemical shift dispersion. The data presented suggest that a full structure determination is possible using the protocol that we have established in the past for the structure determination of the C-terminal domain of presenilin and of Proteorhodopsin.

脂蛋白信号肽酶II（LspA）是位于细菌膜上的细菌蛋白酶。在前脂蛋白穿过内膜输出后，LspA从前脂蛋白切割信号肽。LspA属于一类不寻常的天冬氨酸蛋白酶，在古细菌或真核生物中不存在同源蛋白，这使其成为开发新抗生素的极好靶标。该蛋白质含有164个氨基酸，具有四个预测的α-螺旋。我们希望通过液态NMR光谱确定LspA的结构，并研究环肽抑制剂球霉素如何与蛋白质相互作用。我们已经建立了表达和标记协议，使用我们的无细胞表达系统，并测量了二维和三维NMR光谱的LspA在纳米盘，显示非常好的化学位移分散。所提出的数据表明，一个完整的结构测定是可能的，使用的协议，我们已经建立了在过去的C-末端结构域的早老素和Prorems视紫红质的结构测定。

项目成果

Analyzing native membrane protein assembly in nanodiscs by combined non-covalent mass spectrometry and synthetic biology

• DOI: 10.7554/elife.20954
• 发表时间: 2017-01-09
• 期刊: ELIFE
• 影响因子: 7.7
• 作者: Henrich, Erik;Peetz, Oliver;Morgner, Nini
• 通讯作者: Morgner, Nini