Dannatt Hugh R W, Taylor Garrick F, Varga Krisztina, Higman Victoria A, Pfeil Marc-Philipp, Asilmovska Lubica, Judge Peter J, Watts Anthony
Department of Biochemistry, University of Oxford, South Parks Road, Oxford, OX1 3QU, UK.
J Biomol NMR. 2015 May;62(1):17-23. doi: 10.1007/s10858-015-9911-1. Epub 2015 Feb 21.
We demonstrate that (13)C-detected spectra recorded using fast (60 kHz) magic angle spinning on sub-milligram (<10 μmol) quantities of a protonated 7 trans-membrane helix protein (bacteriorhodopsin) in its native lipid environment are comparable in sensitivity and resolution to those recorded using 15-fold larger sample volumes with conventional solid state NMR methodology. We demonstrate the utility of proton-detected measurements which yield narrow (1)H linewidths under these conditions, and that no structural alterations are observed. We propose that these methods will prove useful to gain structural information on membrane proteins with poor availability, which can be studied in their native lipid environments.
我们证明，在天然脂质环境中，使用快速（60 kHz）魔角旋转对亚毫克量（<10 μmol）的质子化7跨膜螺旋蛋白（细菌视紫红质）记录的¹³C检测光谱，其灵敏度和分辨率与使用传统固态核磁共振方法、样品体积大15倍时记录的光谱相当。我们证明了质子检测测量的实用性，即在这些条件下可产生窄的¹H线宽，并且未观察到结构改变。我们提出，这些方法将被证明对于获取难以获得的膜蛋白的结构信息很有用，这些膜蛋白可以在其天然脂质环境中进行研究。
