Dynamic nuclear polarization-enhanced solid-state NMR of a 13C-labeled signal peptide bound to lipid-reconstituted Sec translocon.

Dynamic nuclear polarization-enhanced solid-state NMR of a 13C-labeled signal peptide bound to lipid-reconstituted Sec translocon.
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与脂质重构的 Sec 易位子结合的 13C 标记信号肽的动态核极化增强固态 NMR。

DOI:
10.1021/ja209378h
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发表时间:
2011
影响因子:
15
通讯作者:
Reggie L
Reggie L
中科院分区:
化学1区
文献类型:
--
作者:
Reggie L

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动态核极化(DNP)使记录结合到脂质重组的SecYEG转位蛋白复合体上的信号肽的二维双量子过滤(DQF)固态核磁共振(ss核磁共振)谱成为可能。样品中的少量多肽(∼40nmol)通常会阻碍多维单链核磁共振实验。如此少量的样本也不例外,因为对于涉及膜蛋白的样本,有限的样本空间大部分被脂类占据。因此,使用这里使用的样品的常规2D DQF单核磁共振谱将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2D光谱。这一进展为膜蛋白的研究开辟了新的可能性,特别是在高分辨率光谱的开发和单个氨基酸信号的指定方面,在这种情况下,信号肽与转位蛋白复合体结合。
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.
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