Breaking symmetry in the structure determination of (large) symmetric protein dimers
Breaking symmetry in the structure determination of (large) symmetric protein dimers
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DOI：
10.1023/a:1020948529076
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发表时间：
2002-10-01
影响因子：
2.7
通讯作者：
Byrd, RA
中科院分区：
文献类型：
--
作者：
Gaponenko, V;Altieri, AS;Byrd, RA
我们展示了一种新的方法来破坏同型二聚体的核磁共振光谱中的对称性。顺磁探针以亚化学计量方式引入，以创建一个不对称体系，顺磁探针仅驻留在二聚体中的一个单体上。这就产生了足够的磁各向异性，可以分辨对称性相关的重叠共振，从而检测每个单体组分特有的伪接触位移和残余偶极耦合。这些假接触位移可以很容易地纳入现有的结构改进计算，并使二聚体蛋白内的单体取向的确定。该方法可广泛用于对称二聚体溶液结构的测定。
We demonstrate a novel methodology to disrupt the symmetry in the NMR spectra of homodimers. A paramagnetic probe is introduced sub-stoichiometrically to create an asymmetric system with the paramagnetic probe residing on only one monomer within the dimer. This creates sufficient magnetic anisotropy for resolution of symmetry-related overlapped resonances and, consequently, detection of pseudocontact shifts and residual dipolar couplings specific to each monomeric component. These pseudocontact shifts can be readily incorporated into existing structure refinement calculations and enable determination of monomer orientation within the dimeric protein. This methodology can be widely used for solution structure determination of symmetric dimers.