Solid-State NMR Derived Structure: Backbone of Influenza A M2 Protein

CrossMCB 9986036这项研究的目的是开发一种新的方法，通过固体核磁共振来表征膜蛋白质的结构，而不需要进行共振指定。这是由于最近在PISEMA(15N-1H偶极-15N化学位移)谱中观察到的共振模式。通过15N均一标记和氨基酸特异性标记相结合，可以获得跨膜螺旋的倾斜和旋转方向。虽然这对于获得早期结构图像来说是一个令人兴奋的发展，但距离和指定的取向约束的组合对于从固体核磁共振衍生的约束中收集全部潜力仍然是重要的。螺旋和螺旋束的详细数学分析有助于对这些约束的最佳利用。通过对所有结构约束和CHARMM能量的改进，有可能获得非常高分辨率的结构。交叉验证将被用来评估这些改进的实验和能量参数中的相对权重。

CrossMCB 9986036The objective of this research is to develop a novel approach to membrane protein structural characterization by solid state NMR without the need for resonance assignments. This is due to the recent observation of resonance patterns in PISEMA (15N-1H dipolar - 15N chemical shift) spectra. By a combination of uniform 15N labeling and amino acid specific labeling, both the tilt and rotational orientation of transmembrane helices can be obtained. While this represents an exciting development for obtaining an early structural image, a combination of distance and assigned orientational constraints will still be important for gleaning the full potential from the solid state NMR-derived constraints. The optimal utilization of these constraints is being aided by a detailed mathematical analysis of helices and helical bundles. Through refinement against all of the structural constraints and the CHARMM energy, it is possible to achieve a very high resolution structure. Cross-validation will be used to assess the relative weights in the experimental and energetic parameters for these refinements.