Structure of amantadine-bound M2 transmembrane peptide of influenza A in lipid bilayers from magic-angle-spinning solid-state NMR: the role of Ser31 in amantadine binding.
Structure of amantadine-bound M2 transmembrane peptide of influenza A in lipid bilayers from magic-angle-spinning solid-state NMR: the role of Ser31 in amantadine binding.
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DOI：
10.1016/j.jmb.2008.11.022
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发表时间：
2009-01-30
影响因子：
5.6
通讯作者：
Hong, Me
中科院分区：
文献类型：
--
作者：
Cady, Sarah D.;Mishanina, Tatiana V.;Hong, Me
甲型流感的M2质子通道是抗病毒药物金刚烷胺和金刚乙胺的靶标，其有效性已被蛋白质跨膜区Ser31到ASN的单点突变所消除。最近从溶液和晶体环境中的洗涤剂增溶蛋白获得的M2跨膜区的高分辨结构给出了相互冲突的药物结合部位。我们给出了Ser31和结合在脂质双层上的M2跨膜肽(M2TMP)中其他一些残基的魔角旋转固体核磁共振结果。比较膜结合的载脂蛋白和络合物M2TMP的光谱表明，Ser31是金刚烷胺最大的化学位移扰动部位。化学位移的限制导致单体结构在Gly34处螺旋轴有一个小的扭结。然后使用之前在无取向双层样品上测量的螺旋倾斜角和几个螺旋间距来构建四聚体模型。这个四聚体模型与溶液和晶体结构的不同之处在于通道N端的开放、Ser31的收缩和Trp41的侧链构象，Trp41是通道门控的重要残基。此外，四聚体模型表明Ser31可能通过氢键与金刚烷胺相互作用。虽然apo和药物结合的M2TMP具有相似的平均结构，但复合肽在生理温度下的线宽要窄得多，这表明药物诱导的膜蛋白动力学发生了变化。此外，在低温下，几个残基在复合肽中显示的线条比载脂蛋白肽更窄，表明金刚烷胺结合降低了特定残基的构象异质性。双层结合的M2TMP的当前固体核磁共振结构与洗涤剂为基础的M2结构的差异表明M2构象对环境敏感，在解释非双层样品的结构发现时必须小心。
The M2 proton channel of influenza A is the target of the antiviral drugs amantadine and rimantadine, whose effectiveness has been abolished by a single-site mutation of Ser31 to Asn in the transmembrane domain of the protein. Recent high-resolution structures of the M2 transmembrane domain obtained from detergent-solubilized protein in solution and crystal environments gave conflicting drug binding sites. We present magic-angle-spinning solid-state NMR results of Ser31 and a number of other residues in the M2 transmembrane peptide (M2TMP) bound to lipid bilayers. Comparison of the spectra of the membrane-bound apo and complexed M2TMP indicates that Ser31 is the site of the largest chemical shift perturbation by amantadine. The chemical shift constraints lead to a monomer structure with a small kink of the helical axis at Gly34. A tetramer model is then constructed using the helix tilt angle and several interhelical distances previously measured on unoriented bilayer samples. This tetramer model differs from the solution and crystal structures in terms of the openness of the N-terminus of the channel, the constriction at Ser31, and the sidechain conformations of Trp41, a residue important for channel gating. Moreover, the tetramer model suggests that Ser31 may interact with amantadine amine via hydrogen bonding. While the apo and drug-bound M2TMP have similar average structures, the complexed peptide has much narrower linewidths at physiological temperature, indicating drug-induced changes of the protein dynamics in the membrane. Further, at low temperature, several residues show narrower lines in the complexed peptide than the apo peptide, indicating that amantadine binding reduces the conformational heterogeneity of specific residues. The differences of the current solid-state NMR structure of the bilayer-bound M2TMP from the detergent-based M2 structures suggest that the M2 conformation is sensitive to the environment, and care must be taken when interpreting structural findings from non-bilayer samples.
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影响因子：
3.4
作者：
Javadpour, MM;Eilers, M;Smith, SO
通讯作者：
Smith, SO
影响因子：
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作者：
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作者：
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DOI：
10.1073/pnas.0804958105
发表时间：
2008-08-05
影响因子：
11.1
作者：
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通讯作者：
Lamb, Robert A.
影响因子：
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作者：
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通讯作者：
COHEN, FE