Franken P, Arold S, Padilla A, Bodeus M, Hoh F, Strub M P, Boyer M, Jullien M, Benarous R, Dumas C
Centre de Biochimie Structurale, UMR C9955 CNRS, U414 INSERM, Université Montpellier I, Faculté de Pharmacie, France.
Protein Sci. 1997 Dec;6(12):2681-3. doi: 10.1002/pro.5560061227.
Human immunodeficiency virus Nef protein accelerates virulent progression of AIDS by its interaction with specific cellular proteins involved in cellular activation and signal transduction. Here we report the purification and crystallization of the conserved core of HIV-1LAI Nef protein in the unliganded form and in complex with the wild-type SH3 domain of the P59fyn protein-tyrosine kinase. One-dimensional NMR experiments show that full-length protein and truncated fragment corresponding to the product of HIV-1 protease cleavage have a well-folded compact tertiary structure. The ligand-free HIV-1 Nefcore protein forms cubic crystals belonging to space group P23 with unit cell dimensions of a = b = c = 86.4 A. The Nef-Fyn SH3 cocrystals belong to the space group P6(1)22 or its enantiomorph, P6(5)22, with unit cell dimensions of a = b = 108.2 A and c = 223.7 A. Both crystal forms diffract to a resolution limit of 3.0 A resolution using synchrotron radiation, and are thus suitable for X-ray structure determination.
人类免疫缺陷病毒Nef蛋白通过与参与细胞活化和信号转导的特定细胞蛋白相互作用，加速艾滋病的恶性进展。在此我们报告了未结合配体形式以及与P59fyn蛋白酪氨酸激酶的野生型SH3结构域形成复合物的HIV-1LAI Nef蛋白保守核心的纯化及结晶情况。一维核磁共振实验表明，全长蛋白以及对应于HIV-1蛋白酶切割产物的截短片段具有折叠良好的紧密三级结构。无配体的HIV-1 Nef核心蛋白形成属于空间群P23的立方晶体，晶胞参数为a = b = c = 86.4 Å。Nef-Fyn SH3共晶体属于空间群P6(1)22或其对映体P6(5)22，晶胞参数为a = b = 108.2 Å，c = 223.7 Å。两种晶体形式利用同步辐射均能衍射到3.0 Å的分辨率极限，因此适合进行X射线结构测定。
