Real-time pure shift ¹⁵N HSQC of proteins: a real improvement in resolution and sensitivity.
Real-time pure shift ¹⁵N HSQC of proteins: a real improvement in resolution and sensitivity.
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DOI：
10.1007/s10858-015-9913-z
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发表时间：
2015-05
影响因子：
2.7
通讯作者：
Morris, Gareth A.
中科院分区：
文献类型：
--
作者：
Kiraly, Peter;Adams, Ralph W.;Paudel, Liladhar;Foroozandeh, Mohammadali;Aguilar, Juan A.;Timari, Istvan;Cliff, Matthew J.;Nilsson, Mathias;Sandor, Peter;Batta, Gyula;Waltho, Jonathan P.;Koever, Katalin E.;Morris, Gareth A.
在质子核磁共振光谱中，由于同核标量耦合的影响，共振分裂成多态，从而降低了光谱分辨率。虽然这些影响往往被隐藏在蛋白质核磁共振波谱低数字分辨率和常规apodization，在幕后同核标量耦合增加光谱拥挤。新的纯位移核磁共振方法为生物分子核磁共振提供了可能性。在不增加实验时间的前提下，提高了分辨率和灵敏度。在这些实验中，自由感应衰减是在短时间的数据采集中收集到的，其持续时间在j -演化的时间尺度上很短，并散布着适当的重聚焦元素。净效应是实时（t 2）宽带同去耦，抑制由质子-质子相互作用引起的多重结构。重聚焦元件的关键特征是它们区分主动（被观察到）和被动（耦合伙伴）自旋的共振。这可以通过使用带选择性重聚焦或BIRD元件来实现，在这两种情况下都伴随着非选择性180°质子脉冲。后一种方法是根据它们与15N的单键异核j耦合选择活性自旋，而前一种方法是选择1H谱的一个区域。提出了几种新的纯位移实验，并对它们提供的代表性样品的分辨率和灵敏度进行了评估：PGK的n端结构域；泛素;以及两个小抗真菌蛋白PAF突变体。这些新的实验，提供更高的灵敏度和分辨率，有可能取代目前的标准HSQC实验。本文的在线版本（doi:10.1007/s10858-015-9913-z）包含补充材料，仅供授权用户使用。
Spectral resolution in proton NMR spectroscopy is reduced by the splitting of resonances into multiplets due to the effect of homonuclear scalar couplings. Although these effects are often hidden in protein NMR spectroscopy by low digital resolution and routine apodization, behind the scenes homonuclear scalar couplings increase spectral overcrowding. The possibilities for biomolecular NMR offered by new pure shift NMR methods are illustrated here. Both resolution and sensitivity are improved, without any increase in experiment time. In these experiments, free induction decays are collected in short bursts of data acquisition, with durations short on the timescale of J-evolution, interspersed with suitable refocusing elements. The net effect is real-time (t 2) broadband homodecoupling, suppressing the multiplet structure caused by proton–proton interactions. The key feature of the refocusing elements is that they discriminate between the resonances of active (observed) and passive (coupling partner) spins. This can be achieved either by using band-selective refocusing or by the BIRD element, in both cases accompanied by a nonselective 180° proton pulse. The latter method selects the active spins based on their one-bond heteronuclear J-coupling to 15N, while the former selects a region of the 1H spectrum. Several novel pure shift experiments are presented, and the improvements in resolution and sensitivity they provide are evaluated for representative samples: the N-terminal domain of PGK; ubiquitin; and two mutants of the small antifungal protein PAF. These new experiments, delivering improved sensitivity and resolution, have the potential to replace the current standard HSQC experiments. The online version of this article (doi:10.1007/s10858-015-9913-z) contains supplementary material, which is available to authorized users.
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影响因子：
16.6
作者：
Meyer, N. Helge;Zangger, Klaus
通讯作者：
Zangger, Klaus
DOI：
10.1006/jmra.1994.1227
发表时间：
1994-11-01
期刊：
JOURNAL OF MAGNETIC RESONANCE SERIES A
影响因子：
--
作者：
KONTAXIS, G;STONEHOUSE, J;KEELER, J
通讯作者：
KEELER, J
DOI：
10.1111/j.1742-4658.2009.07011.x
发表时间：
2009-05
期刊：
The FEBS journal
影响因子：
--
作者：
Batta G;Barna T;Gáspári Z;Sándor S;Kövér KE;Binder U;Sarg B;Kaiserer L;Chhillar AK;Eigentler A;Leiter E;Hegedüs N;Pócsi I;Lindner H;Marx F
通讯作者：
Marx F
影响因子：
2.2
作者：
Lupulescu, Adonis;Olsen, Gregory L.;Frydman, Lucio
通讯作者：
Frydman, Lucio
影响因子：
2.9
作者：
Meyer, N. Helge;Zangger, Klaus
通讯作者：
Zangger, Klaus