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）是位于细菌膜中的细菌蛋白酶。 LSPA在整个膜上导出后，信号肽从刺激蛋白中切开。 LSPA属于一类不寻常的天冬氨酸蛋白酶，该蛋白酶在古细菌或真核生物中没有同源蛋白质，这使其成为开发新抗生素的绝佳靶标。该蛋白质含有164个氨基酸，带有四个预测的α-螺旋。我们希望通过液态NMR光谱确定LSPA的结构，并研究环状肽抑制剂球霉素如何与蛋白质相互作用。我们已经使用我们的无细胞表达系统建立了表达和标记方案，并在纳米散发中测量了LSPA的2D和3D NMR光谱，这些纳米盘表现出非常好的化学位移分散体。提供的数据表明，使用我们过去在过去的结构确定Presenilin和Proteorhopopsin的C末端结构域的结构确定的情况下，可以进行完整的结构确定。

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