M2 proton channel structural validation from full-length protein samples in synthetic bilayers and E. coli membranes.
M2 proton channel structural validation from full-length protein samples in synthetic bilayers and E. coli membranes.
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DOI：
10.1002/anie.201204666
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发表时间：
2012-08-13
影响因子：
16.6
通讯作者：
Cross, Timothy A.
中科院分区：
文献类型：
--
作者：
Miao, Yimin;Qin, Huajun;Fu, Riqiang;Sharma, Mukesh;Can, Thach V.;Hung, Ivan;Luca, Sorin;Gor'kov, Peter L.;Brey, William W.;Cross, Timothy A.
膜蛋白的结构和功能，特别是对于小的膜蛋白，可以对用于结构表征的膜模拟环境高度敏感，如来自甲型流感病毒的M2蛋白所例示的，其已经在液晶脂质双层、洗涤剂胶束和基于洗涤剂的晶体中表征。[3-8]各种跨膜（TM）螺旋倾斜角，不同的药物结合位点和两亲性螺旋相互作用，以及缺乏共识的侧链几何形状的功能关键残基是从这组结构是显而易见的。这些结构差异中的许多可以根据蛋白质环境的影响来解释。疏水厚度影响螺旋倾斜;洗涤剂渗透到螺旋束和晶体接触影响包装，因此倾斜的螺旋，而胶束的高度弯曲的表面不稳定的两亲性螺旋与什么将是双层界面的相互作用。[9]这些结构扰动可以影响功能特性，例如抗病毒药物与蛋白质的结合以及我们对质子通道功能机制的理解。在这里探索了天然膜需要建模以实现天然膜蛋白结构的准确程度，我们的目标是验证四聚体M2电导结构域（M2 CD;残基22-62; PDB#2L 0 J）的结构，该结构已在合成脂质双层中进行了结构表征。我们已经着手通过观察合成双层中的全长蛋白质来做到这一点，以及在天然E。coli膜。我们首次报道了使用魔角自旋固态NMR（ssNMR）从全长M2（M2 FL）蛋白质中获得的结构见解，并在将其插入E.大肠杆菌膜的细胞器，而从来没有暴露于洗涤剂的环境。这些结果验证了早期的结构结果从M2 CD中观察到的液晶双层envelope.In除了它的质子通道活性，M2促进病毒出芽，本地化的蛋白质在这个网站上的感染细胞的表面上表明，M2是不是嵌入在高胆固醇和鞘磷脂的环境中占主导地位的病毒颗粒。[10相反，M2似乎优先定位于筏状脂质结构域的外周，病毒颗粒从筏状脂质结构域出芽，并且M2的两亲性螺旋
Membrane protein structure and function, especially for small membrane proteins, can be highly sensitive to the membrane mimetic environment used for structural characterization, as exemplified by the M2 protein from influenza A virus that has been characterized in liquid crystalline lipid bilayers, detergent micelles and in detergent based crystals.[3–8] Various transmembrane (TM) helical tilt angles, different drug binding sites and amphipathic helix interactions, as well as a lack of consensus on the sidechain geometry for the functionally critical residues is apparent from this set of structures. Many of these structural differences can be explained based on the influence of the protein's environment. Hydrophobic thickness influences the helical tilt; detergent penetration into the helical bundle and crystal contacts influence the packing and hence tilt of the helices, while the highly curved surface of micelles destabilize the interactions of amphipathic helices with what would be the bilayer interface.[9] These structural perturbations can influence functional properties such as the binding of the antiviral drug to the protein and our understanding of the proton channel functional mechanism. Exactly how well the native membrane needs to be modeled to achieve a native membrane protein structure is explored here, where we aim to validate the structure of the tetrameric M2 conductance domain (M2CD; residues 22–62; PDB# 2L0J) that has been structurally characterized in synthetic lipid bilayers. We have set out to do this by observing the full length protein in synthetic bilayers, as well as in native E. coli membranes. For the first time we report on structural insights from the full length M2 (M2FL) protein using magic angle spinning solid state NMR (ssNMR) and we present spectra of the protein as it is inserted into the E. coli membranes by the cellular apparatus without ever being exposed to a detergent environment. These results validate the earlier structural results obtained from the M2CD observed in a liquid crystalline bilayer envionment.In addition to its proton channel activity, M2 facilitates viral budding that localizes the protein to this site on the surface of an infected cell suggesting that M2 is not imbedded in the high cholesterol and sphingomyelin environment that dominates the viral particle.[10, 11] Instead, M2 appears to be preferentially localized to the periphery of the raft-like lipid domains from which the viral particle buds and where the amphipathic helices of the M2
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影响因子：
3.7
作者：
Rossman JS;Lamb RA
通讯作者：
Lamb RA
影响因子：
64.5
作者：
LAMB, RA;ZEBEDEE, SL;RICHARDSON, CD
通讯作者：
RICHARDSON, CD
影响因子：
4.8
作者：
Tang, YJ;Zaitseva, F;Pinto, LH
通讯作者：
Pinto, LH
影响因子：
3
作者：
Kamihira, M;Vosegaard, T;Watts, A
通讯作者：
Watts, A
来自固态NMR的流感M2质子通道中质子传导和门控的机制。
DOI：
10.1126/science.1191714
发表时间：
2010-10-22
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Hu F;Luo W;Hong M
通讯作者：
Hong M