Improving the diffraction of full-length human selenomethionyl metavinculin crystals by streak-seeding.
Improving the diffraction of full-length human selenomethionyl metavinculin crystals by streak-seeding.
复制标题
通过划线播种改善全长人硒代甲硫氨酰美维库林晶体的衍射。
DOI：
10.1107/s1744309110041059
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
Izard,Tina
中科院分区：
文献类型：
--
作者：
Rangarajan,ErumbiS;Izard,Tina
Metavinculin是一种选择性剪接的vinculin异构体，在其尾部区域有68个残基插入（总共1134个残基），并且只在心脏和平滑肌组织中表达，在心肌细胞粘附复合物中起重要作用。metavinculin特异性插入的突变与男性扩张型心肌病（DCM）有关。悬垂气相扩散生长的全长硒代蛋氨酸标记的metavinculin突变体的dcm相关晶体衍射差，对辐射高度敏感，无法以最高分辨率收集完整的x射线衍射数据集。条纹播种显著提高了dcm相关突变体metavinculin晶体的稳定性、晶体生长速度和衍射质量，使完整的数据收集达到3.9 Å分辨率。这些晶体属于空间群P43212，两个分子在不对称单元和单元胞参数a = b = 170, c = 211 Å， α = β = γ = 90°。
Metavinculin is an alternatively spliced isoform of vinculin that has a 68-residue insert in its tail domain (1134 total residues) and is exclusively expressed in cardiac and smooth muscle tissue, where it plays important roles in myocyte adhesion complexes. Mutations in the metavinculin-specific insert are associated with dilated cardiomyopathy (DCM) in man. Crystals of a DCM-associated mutant of full-length selenomethionine-labeled metavinculin grown by hanging-drop vapor diffusion diffracted poorly and were highly sensitive to radiation, preventing the collection of a complete X-ray diffraction data set at the highest possible resolution. Streak-seeding markedly improved the stability, crystal-growth rate and diffraction quality of DCM-associated mutant metavinculin crystals, allowing complete data collection to 3.9 Å resolution. These crystals belonged to space group P43212, with two molecules in the asymmetric unit and unit-cell parameters a = b = 170, c = 211 Å, α = β = γ = 90°.