Structure and dynamics of a membrane protein in micelles from three solution NMR experiments
Structure and dynamics of a membrane protein in micelles from three solution NMR experiments
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DOI：
10.1023/a:1024047805043
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发表时间：
2003-08-01
影响因子：
2.7
通讯作者：
Opella, SJ
中科院分区：
文献类型：
--
作者：
Lee, S;Mesleh, MF;Opella, SJ
三个解决方案的NMR实验上均匀N-15标记的膜蛋白胶束提供了足够的信息来描述其螺旋的结构，拓扑结构和动力学，以及额外的信息，其特征在于在终端和螺旋间环区域的残基的主要功能。骨架酰胺共振通过HMQC-NOESY实验进行分配，骨架动力学通过H-1-N-15杂环NOE实验进行表征，该实验明确区分了蛋白质末端和螺旋间环区域中的结构化螺旋残基和更多移动的残基。螺旋的结构和拓扑结构由偶极波和比萨轮描述，这些轮来自于残余偶极耦合（RDC）和残余化学位移各向异性（RCSA）的实验测量。结果表明，膜结合型Pf 1外壳蛋白具有20个残基的跨膜疏水螺旋，其取向与8个残基的两亲螺旋相差约90度。这三个实验的组合，产生偶极波和比萨轮有可能有助于膜蛋白的高通量结构表征。
Three solution NMR experiments on a uniformly N-15 labeled membrane protein in micelles provide sufficient information to describe the structure, topology, and dynamics of its helices, as well as additional information that characterizes the principal features of residues in terminal and inter-helical loop regions. The backbone amide resonances are assigned with an HMQC-NOESY experiment and the backbone dynamics are characterized by a H-1-N-15 heteronuclear NOE experiment, which clearly distinguishes between the structured helical residues and the more mobile residues in the terminal and inter-helical loop regions of the protein. The structure and topology of the helices are described by Dipolar waves and PISA wheels derived from experimental measurements of residual dipolar couplings (RDCs) and residual chemical shift anisotropies (RCSAs). The results show that the membrane-bound form of Pf1 coat protein has a 20-residue trans-membrane hydrophobic helix with an orientation that differs by about 90degrees from that of an 8-residue amphipathic helix. This combination of three-experiments that yields Dipolar waves and PISA wheels has the potential to contribute to high-throughput structural characterizations of membrane proteins.