Okemefuna Azubuike I, Gilbert Hannah E, Griggs Kim M, Ormsby Rebecca J, Gordon David L, Perkins Stephen J
Department of Biochemistry and Molecular Biology, Darwin Building, University College London, Gower Street, London WC1E 6BT, UK.
J Mol Biol. 2008 Jan 4;375(1):80-101. doi: 10.1016/j.jmb.2007.09.026. Epub 2007 Sep 14.
Factor H (FH) is a plasma glycoprotein that plays a central role in regulation of the alternative pathway of complement. It is composed of 20 short complement regulator (SCR) domains. The SCR-1/5 fragment is required for decay acceleration and cofactor activity, while the SCR-16/20 fragment possesses binding sites for complement C3d and heparin. X-ray scattering and analytical ultracentrifugation showed that SCR-1/5 was monomeric, while SCR-16/20 formed dimers. The Guinier radius of gyration R(G) of 4.3 nm for SCR-1/5 and those of 4.7 nm and about 7.8 nm for monomeric and dimeric SCR-16/20, respectively, showed that their structures are partially folded back and bent. The distance distribution function P(r) showed that SCR-1/5 has a maximum dimension of 15 nm while monomeric and dimeric SCR-16/20 are 17 nm and about 27 nm long, respectively. The sedimentation coefficient of 2.4 S for SCR-1/5 showed no concentration-dependence, while that for SCR-16/20 was 2.8 S for the monomer and 3.9 S for the dimer. Sedimentation equilibrium data showed that SCR-1/5 is monomeric while SCR-16/20 exhibited a weak monomer-dimer equilibrium with a dissociation constant of 16 microM. The constrained scattering and sedimentation modelling of SCR-1/5 and SCR-16/20 showed that partially folded-back and bent flexible SCR arrangements fitted both data sets better than extended linear arrangements, and that the dimer was best modelled in the SCR-16/20 model by an end-to-end association of two SCR-20 domains. The SCR-1/5 and SCR-16/20 models were conformationally similar to the previously determined partially folded-back structure for intact wild-type FH, hence suggesting a partial explanation of the intact FH structure. Comparison of the SCR-16/20 model with the crystal structure of C3b clarified reasons for the distribution of mutations leading to atypical haemolytic uraemic syndrome.
补体因子H(FH)是一种血浆糖蛋白,在补体替代途径的调节中起核心作用。它由20个短补体调节蛋白(SCR)结构域组成。SCR-1/5片段是衰变加速和辅因子活性所必需的,而SCR-16/20片段具有补体C3d和肝素的结合位点。X射线散射和分析超速离心表明,SCR-1/5是单体,而SCR-16/20形成二聚体。SCR-1/5的吉尼埃回转半径R(G)为4.3nm,单体和二聚体SCR-16/20的回转半径分别为4.7nm和约7.8nm,这表明它们的结构部分折叠并弯曲。距离分布函数P(r)表明,SCR-1/5的最大尺寸为15nm,而单体和二聚体SCR-16/20的长度分别为17nm和约27nm。SCR-1/5的沉降系数为2.4S,不依赖于浓度,而SCR-16/20的单体沉降系数为2.8S,二聚体沉降系数为3.9S。沉降平衡数据表明,SCR-1/5是单体,而SCR-16/20表现出弱的单体-二聚体平衡,解离常数为16μM。SCR-1/5和SCR-16/20的受限散射和沉降建模表明,部分折叠和弯曲的柔性SCR排列比延伸的线性排列更能拟合这两组数据,并且在SCR-16/20模型中,二聚体最好通过两个SCR-20结构域的端对端缔合来建模。SCR-1/5和SCR-16/20模型在构象上与先前确定的完整野生型FH部分折叠的结构相似,因此对完整FH结构提出了部分解释。将SCR-16/20模型与C3b的晶体结构进行比较,阐明了导致非典型溶血性尿毒症综合征的突变分布的原因。