Schmidt Christoph Q, Herbert Andrew P, Mertens Haydyn D T, Guariento Mara, Soares Dinesh C, Uhrin Dusan, Rowe Arthur J, Svergun Dmitri I, Barlow Paul N
Edinburgh Biomolecular NMR Unit, Centre for Chemical and Translational Biology, Schools of Biological Sciences and Chemistry, University of Edinburgh, Edinburgh EH9 3JJ, UK.
J Mol Biol. 2010 Jan 8;395(1):105-22. doi: 10.1016/j.jmb.2009.10.010. Epub 2009 Oct 14.
The first eight and the last two of 20 complement control protein (CCP) modules within complement factor H (fH) encompass binding sites for C3b and polyanionic carbohydrates. These binding sites cooperate self-surface selectively to prevent C3b amplification, thus minimising complement-mediated damage to host. Intervening fH CCPs, apparently devoid of such recognition sites, are proposed to play a structural role. One suggestion is that the generally small CCPs 10-15, connected by longer-than-average linkers, act as a flexible tether between the two functional ends of fH; another is that the long linkers induce a 180 degrees bend in the middle of fH. To test these hypotheses, we determined the NMR-derived structure of fH12-13 consisting of module 12, shown here to have an archetypal CCP structure, and module 13, which is uniquely short and features a laterally protruding helix-like insertion that contributes to a prominent electropositive patch. The unusually long fH12-13 linker is not flexible. It packs between the two CCPs that are not folded back on each other but form a shallow vee shape; analytical ultracentrifugation and X-ray scattering supported this finding. These two techniques additionally indicate that flanking modules (within fH11-14 and fH10-15) are at least as rigid and tilted relative to neighbours as are CCPs 12 and 13 with respect to one another. Tilts between successive modules are not unidirectional; their principal axes trace a zigzag path. In one of two arrangements for CCPs 10-15 that fit well with scattering data, CCP 14 is folded back onto CCP 13. In conclusion, fH10-15 forms neither a flexible tether nor a smooth bend. Rather, it is compact and has embedded within it a CCP module (CCP 13) that appears to be highly specialised given both its deviant structure and its striking surface charge distribution. A passive, purely structural role for this central portion of fH is unlikely.
补体因子H(fH)中的20个补体控制蛋白(CCP)模块的前8个和后2个包含C3b和多阴离子碳水化合物的结合位点。这些结合位点选择性地协同作用于自身表面,以防止C3b的扩增,从而将补体介导的对宿主的损伤降至最低。中间的fH CCPs显然没有此类识别位点,推测其发挥结构作用。一种观点认为,通常较小的CCP 10 - 15由比平均长度更长的连接子相连,在fH的两个功能端之间起到柔性系链的作用;另一种观点是,长连接子会使fH在中间形成180度的弯曲。为了验证这些假设,我们确定了由模块12和模块13组成的fH12 - 13的核磁共振衍生结构,此处显示模块12具有典型的CCP结构,而模块13独特地短小,具有侧向突出的螺旋状插入结构,形成一个显著的正电斑块。异常长的fH12 - 13连接子并不灵活。它堆积在两个未相互折叠而是形成浅V形的CCP之间;分析型超速离心和X射线散射支持了这一发现。这两种技术还表明,侧翼模块(在fH11 - 14和fH10 - 15内)至少与CCP 12和13相对于彼此一样刚性且相对于相邻模块倾斜。连续模块之间的倾斜并非单向的;它们的主轴呈锯齿状路径。在与散射数据拟合良好的CCP 10 - 15的两种排列之一中,CCP 14折叠在CCP 13上。总之,fH10 - 15既不形成柔性系链也不形成平滑弯曲。相反,它结构紧凑,其中嵌入了一个CCP模块(CCP 13),鉴于其异常的结构和显著的表面电荷分布,该模块似乎高度特化。fH这一中心部分不太可能仅起被动的纯粹结构作用。