Cai M, Huang Y, Ghirlando R, Wilson K L, Craigie R, Clore G M
Laboratory of Chemical Physics, Building 5, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892-0510, USA.
EMBO J. 2001 Aug 15;20(16):4399-407. doi: 10.1093/emboj/20.16.4399.
The nuclear envelope proteins LAP2, emerin and MAN1 share a conserved approximately 40-residue 'LEM' motif. Loss of emerin causes Emery-Dreifuss muscular dystrophy. We have solved the solution NMR structure of the constant region of human LAP2 (residues 1-168). Human LAP2(1-168) has two structurally independent, non-interacting domains located at residues 1-50 ('LAP2-N') and residues 111-152 (LEM-domain), connected by an approximately 60-residue flexible linker. The two domains are structurally homologous, comprising a helical turn followed by two helices connected by an 11-12-residue loop. This motif is shared by subdomains of T4 endonuclease VII and transcription factor rho, despite negligible (< or =15%) sequence identity. NMR chemical shift mapping demonstrated that the LEM-domain binds BAF (barrier-to-autointegration factor), whereas LAP2-N binds DNA. Both binding surfaces comprise helix 1, the N-terminus of helix 2 and the inter-helical loop. Binding selectivity is determined by the nature of the surface residues in these binding sites, which are predominantly positively charged for LAP2-N and hydrophobic for the LEM-domain. Thus, LEM and LEM-like motifs form a common structure that evolution has customized for binding to BAF or DNA.
核被膜蛋白LAP2、emerin和MAN1共享一个保守的约40个残基的“LEM”基序。emerin的缺失会导致埃默里-德赖富斯肌营养不良症。我们已经解析了人LAP2恒定区(1-168位残基)的溶液核磁共振结构。人LAP2(1-168)有两个结构上独立、不相互作用的结构域,分别位于1-50位残基(“LAP2-N”)和111-152位残基(LEM结构域),由一个约60个残基的柔性连接子相连。这两个结构域在结构上同源,包括一个螺旋转角,随后是由一个11-12个残基的环连接的两个螺旋。尽管序列同一性可忽略不计(≤15%),但该基序为T4内切核酸酶VII和转录因子rho的亚结构域所共有。核磁共振化学位移图谱表明,LEM结构域结合BAF(屏障自整合因子),而LAP2-N结合DNA。两个结合表面均包括螺旋1、螺旋2的N端和螺旋间环。结合选择性由这些结合位点表面残基的性质决定,LAP2-N的表面残基主要带正电荷,而LEM结构域的表面残基为疏水性。因此,LEM和类LEM基序形成了一种共同结构,进化已对其进行定制以结合BAF或DNA。