Department of Chemistry, University of Hull, Hull HU6 7RX, United Kingdom.
J Am Chem Soc. 2011 Nov 30;133(47):19084-6. doi: 10.1021/ja209378h. Epub 2011 Nov 7.
Dynamic nuclear polarization (DNP) has made it possible to record 2D double-quantum-filtered (DQF) solid-state NMR (ssNMR) spectra of a signal peptide bound to a lipid-reconstituted SecYEG translocon complex. The small quantity of peptide in the sample (~40 nmol) normally prohibits multidimensional ssNMR experiments. Such small amounts are not the exception, because for samples involving membrane proteins, most of the limited sample space is occupied by lipids. As a consequence, a conventional 2D DQF ssNMR spectrum with the sample used here would require many weeks if not months of measurement time. With the help of DNP, however, we were able to acquire such a 2D spectrum within 20 h. This development opens up new possibilities for membrane protein studies, particularly in the exploitation of high-resolution spectroscopy and the assignment of individual amino acid signals, in this case for a signal peptide bound to the translocon complex.
动态核极化（DNP）使得记录与脂质重建的 SecYEG 转运通道复合物结合的信号肽的二维双量子过滤（DQF）固态 NMR（ssNMR）谱成为可能。样品中信号肽的数量很少（约 40nmol），通常会禁止多维 ssNMR 实验。这种少量的情况并不罕见，因为对于涉及膜蛋白的样品，大部分有限的样品空间都被脂质占据。因此，如果使用这里的样品，常规的二维 DQF ssNMR 谱需要数周甚至数月的测量时间。然而，在 DNP 的帮助下，我们能够在 20 小时内获得这样的二维谱。这一发展为膜蛋白研究开辟了新的可能性，特别是在利用高分辨率光谱学和分配单个氨基酸信号方面，在这种情况下，就是分配与转运通道复合物结合的信号肽。
