Metalloantibody design.
Metalloantibody design.
复制标题
金属抗体设计。
DOI：
10.1096/fasebj.9.1.7821766
复制
发表时间：
1995
期刊：
影响因子：
--
通讯作者：
Getzoff,ED
中科院分区：
文献类型：
--
作者：
Roberts,VA;Getzoff,ED
基于三维抗体模型和锌结合金属酶的晶体结构，在催化抗体43C9的抗原结合口袋内设计了金属结合位点。这些四面体锌结合位点被设计成模拟酶中催化金属位点的二级和三级结构特征。每个位点都计划在相邻的反平行β -链上有两个he配体。选择位点将金属离子从散装溶剂中分离出来，并在抗原或潜在底物旁边放置一个开放的金属配位。三种不同的金属位点设计，配体在可变轻域，可变重域，以及在两个域，后来被实验实现，并在光谱上显示出与预测的金属离子结合。这些结果证明了我们的设计方法的成功，金属蛋白设计抗体结构的多功能性，以及三维模型的有效性。在口袋底部有序抗原识别区域中可预测地设计多个金属位点的能力允许调整金属离子的放置，并增强与假定底物相互作用的可能性。-Roberts， V. A, Getzoff， E. D.金属抗体设计。中国生物医学工程学报（英文版），2004 - 04 （1995）
Metal‐binding sites were designed within the antigen‐binding pocket of the catalytic antibody 43C9 based on a 3‐dimensional antibody model and crystallographic structures of Zn‐binding metalloenzymes. These tetrahedral Zn‐binding sites were designed to mimic both secondary and tertiary structural characteristics of catalytic metal sites in enzymes. Each site was planned to have two His ligands across from each other on adjacent antiparallel β‐strands. Sites were selected to sequester the metal ion from bulk solvent and place an open metal coordination position next to the antigen or potential substrates. Three distinct metal‐site designs, with ligands in the variable light domain, in the variable heavy domain, and in both domains, were later implemented experimentally and shown spectroscopically to bind metal ions as predicted. These results demonstrate the success of our design approach, the versatility of the antibody structure for metalloprotein design, and the validity of the 3‐dimensional model. The ability to predictably design multiple metal sites in the ordered antigen‐recognition region at the bottom of the pocket allows tuning of metal ion placement and enhances the likelihood of interaction with putative substrates.—Roberts, V. A., Getzoff, E. D. Metalloantibody design.FASEB J.9, 94‐100 (1995)