Agarwal Vipin, Fink Uwe, Schuldiner Shimon, Reif Bernd
Leibniz-Institut für Molekulare Pharmakologie (FMP), Robert-Rössle-Str. 10, D-13125 Berlin, Germany.
Biochim Biophys Acta. 2007 Dec;1768(12):3036-43. doi: 10.1016/j.bbamem.2007.09.012. Epub 2007 Oct 2.
We study the uniformly 13C,15N isotopically enriched Escherichia coli multidrug resistance transporter EmrE using MAS solid-state NMR. Solid-state NMR can provide complementary structural information as the method allows studying membrane proteins in their native environment as no detergent is required for reconstitution. We compare the spectra obtained from wildtype EmrE to those obtained from the mutant EmrE-E14C. To resolve the critical amino acid E14, glutamic/aspartic acid selective experiments are carried out. These experiments allow to assign the chemical shift of the carboxylic carbon of E14. In addition, spectra are analyzed which are obtained in the presence and absence of the ligand TPP+.
我们使用魔角旋转（MAS）固态核磁共振（NMR）技术研究了均匀13C、15N同位素富集的大肠杆菌多药耐药转运蛋白EmrE。固态NMR能够提供互补的结构信息，因为该方法允许在蛋白质的天然环境中研究膜蛋白，重构过程无需去污剂。我们将野生型EmrE的光谱与突变型EmrE-E14C的光谱进行了比较。为了确定关键氨基酸E14，我们进行了谷氨酸/天冬氨酸选择性实验。这些实验能够确定E14羧基碳的化学位移。此外，还分析了在存在和不存在配体TPP+的情况下获得的光谱。
