Complexes of neutralizing and non-neutralizing affinity matured Fabs with a mimetic of the internal trimeric coiled-coil of HIV-1 gp41.
Complexes of neutralizing and non-neutralizing affinity matured Fabs with a mimetic of the internal trimeric coiled-coil of HIV-1 gp41.
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DOI：
10.1371/journal.pone.0078187
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Wlodawer A
中科院分区：
文献类型：
--
作者：
Gustchina E;Li M;Ghirlando R;Schuck P;Louis JM;Pierson J;Rao P;Subramaniam S;Gustchina A;Clore GM;Wlodawer A
先前已构建并报道了一系列针对 HIV-1 gp41 N-七肽重复序列 (N-HR) 的保守内部三聚体卷曲螺旋的微型抗体（单价和二价 Fab）。两个密切相关的单价 Fab 的晶体结构，一个 (Fab 8066) 对多种 HIV-1 亚型 B 和 C 病毒具有广泛中和作用，另一个 (Fab 8062) 具有非中和作用，代表该系列的极端情况，之前已将其作为与 5-Helix（一种 gp41 前发夹中间体模拟物）的复合物进行解析。这些 Fab 与 HIV-1 gp41 N 肽的共价稳定嵌合三聚体（称为 (CCIZN36)3 或 3-H）的结合现已使用 X 射线晶体学、冷冻电子显微镜和各种生物物理方法进行了研究。 3-H 与 Fab 8066 和 Fab 8062 之间的复合物的晶体结构分别以 2.8 和 3.0 Å 的分辨率测定。尽管中和性 Fab 8066 及其非中和性对应物 Fab 8062 的复合物结构大体相似，但它们之间的微小差异可能与这些抗体的生物学特性相关。与3-H和5-Helix形成的复合物中每种抗体相应CDR的构象非常相似。复合物形成时对不同靶标的适应主要是通过 N-HR 螺旋三聚体结构的变化以及通过刚体运动调整复合物中 Fab 分子相对于 N-HR 的方向来实现的。这里提供的结构数据表明，与 Fab 8066 的结合相比，三个 Fab 8062 的高亲和力结合需要 N-HR 三聚体的结构发生更显着的变化。对与不同 gp41 中间模拟物复合的 Fab 结构进行比较分析，可以进一步评估中和抗体生成的生物学相关性，并为免疫原设计提供新的结构见解。
A series of mini-antibodies (monovalent and bivalent Fabs) targeting the conserved internal trimeric coiled-coil of the N-heptad repeat (N-HR) of HIV-1 gp41 has been previously constructed and reported. Crystal structures of two closely related monovalent Fabs, one (Fab 8066) broadly neutralizing across a wide panel of HIV-1 subtype B and C viruses, and the other (Fab 8062) non-neutralizing, representing the extremes of this series, were previously solved as complexes with 5-Helix, a gp41 pre-hairpin intermediate mimetic. Binding of these Fabs to covalently stabilized chimeric trimers of N-peptides of HIV-1 gp41 (named (CCIZN36)3 or 3-H) has now been investigated using X-ray crystallography, cryo-electron microscopy, and a variety of biophysical methods. Crystal structures of the complexes between 3-H and Fab 8066 and Fab 8062 were determined at 2.8 and 3.0 Å resolution, respectively. Although the structures of the complexes with the neutralizing Fab 8066 and its non-neutralizing counterpart Fab 8062 were generally similar, small differences between them could be correlated with the biological properties of these antibodies. The conformations of the corresponding CDRs of each antibody in the complexes with 3-H and 5-Helix are very similar. The adaptation to a different target upon complex formation is predominantly achieved by changes in the structure of the trimer of N-HR helices, as well as by adjustment of the orientation of the Fab molecule relative to the N-HR in the complex, via rigid-body movement. The structural data presented here indicate that binding of three Fabs 8062 with high affinity requires more significant changes in the structure of the N-HR trimer compared to binding of Fab 8066. A comparative analysis of the structures of Fabs complexed to different gp41 intermediate mimetics allows further evaluation of biological relevance for generation of neutralizing antibodies, as well as provides novel structural insights into immunogen design.
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影响因子：
4.8
作者：
Cai, ML;Williams, DC;Clore, GM
通讯作者：
Clore, GM
DOI：
10.1107/s0907444904019158
发表时间：
2004-12-01
影响因子：
2.2
作者：
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通讯作者：
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影响因子：
46.9
作者：
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通讯作者：
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影响因子：
5.4
作者：
Gustchina, Elena;Louis, John M.;Clore, G. Marius
通讯作者：
Clore, G. Marius
影响因子：
6.7
作者：
Gustchina E;Li M;Louis JM;Anderson DE;Lloyd J;Frisch C;Bewley CA;Gustchina A;Wlodawer A;Clore GM
通讯作者：
Clore GM