Lewandowski Józef R, De Paëpe Gaël, Eddy Matthew T, Struppe Jochem, Maas Werner, Griffin Robert G
Department of Chemistry and Francis Bitter Magnet Laboratory, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139, USA.
J Phys Chem B. 2009 Jul 9;113(27):9062-9. doi: 10.1021/jp810280t.
We demonstrate the successful application of (13)C-(13)C proton assisted recoupling (PAR) on [U-(13)C,(15)N] N-f-MLF-OH and [U-(13)C,(15)N] protein GB1 at high magic angle spinning (MAS) frequencies (omega(r)/2pi = 65 kHz). Specifically, by combining PAR mixing with low power heteronuclear decoupling (omega(1H)/2pi approximately 16 kHz) and high spinning frequencies, we obtain high resolution 2D spectra displaying long-range (13)C-(13)C contacts from which distance estimates can be extracted. These experiments therefore demonstrate the possibility of performing high resolution structural studies in the limit of high spinning frequency and low power (1)H decoupling, a regime which optimizes the resolution of protein samples and preserves their integrity.
我们展示了（13）C - （13）C质子辅助重耦合（PAR）在高魔角旋转（MAS）频率（ωr/2π = 65 kHz）下对[U - （13）C，（15）N] N - f - MLF - OH和[U - （13）C，（15）N]蛋白质GB1的成功应用。具体而言，通过将PAR混合与低功率异核去耦（ω1H/2π约为16 kHz）和高旋转频率相结合，我们获得了高分辨率二维光谱，显示了可从中提取距离估计值的远程（13）C - （13）C接触。因此，这些实验证明了在高旋转频率和低功率（1）H去耦的极限条件下进行高分辨率结构研究的可能性，这种条件优化了蛋白质样品的分辨率并保持其完整性。
