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首页> 外文期刊>Journal of Molecular Biology >Crystal Structure of the Anti-His Tag Antibody 3D5 Single-chain Fragment Complexed to its Antigen.
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Crystal Structure of the Anti-His Tag Antibody 3D5 Single-chain Fragment Complexed to its Antigen.

机译：与他的抗原复合的抗His标签抗体3D5单链片段的晶体结构。

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摘要

The crystal structure of a mutant form of the single-chain fragment (scFv), derived from the monoclonal anti-His tag antibody 3D5, in complex with a hexahistidine peptide has been determined at 2.7 A resolution. The peptide binds to a deep pocket formed at the interface of the variable domains of the light and the heavy chain, mainly through hydrophobic interaction to aromatic residues and hydrogen bonds to acidic residues. The antibody recognizes the C-terminal carboxylate group of the peptide as well as the main chain of the last four residues and the last three imidazole side-chains. The crystals have a solvent content of 77% (v/v) and form 70 A-wide channels that would allow the diffusion of peptides or even small proteins. The anti-His scFv crystals could thus act as a framework for the crystallization of His-tagged target proteins. Designed mutations in framework regions of the scFv lead to high-level expression of soluble protein in the periplasm of Escherichia coli. The recombinant anti-His scFv is a convenient detection tool when fused to alkaline phosphatase. When immobilized on a matrix, the antibody can be used for affinity purification of recombinant proteins carrying a very short tag of just three histidine residues, suitable for crystallization. The experimental structure is now the basis for the design of antibodies with even higher stability and affinity. (c) 2002 Elsevier Science Ltd.
机译：已经确定了与六组氨酸肽复合的衍生自单克隆抗His标签抗体3D5的单链片段（scFv）的突变形式的晶体结构。该肽主要通过与芳族残基的疏水相互作用和与酸性残基的氢键结合至形成在轻链和重链可变域界面处的深袋。抗体识别肽的C末端羧酸酯基团，以及最后四个残基的主链和最后三个咪唑侧链。晶体的溶剂含量为77％（v / v），并形成70 A宽的通道，该通道将允许肽甚至小蛋白的扩散。因此，抗His scFv晶体可作为带有His标签的靶蛋白结晶的框架。在scFv的框架区域中设计的突变会导致可溶性蛋白在大肠杆菌的周质中高水平表达。重组抗His scFv与碱性磷酸酶融合时是一种方便的检测工具。当固定在基质上时，该抗体可用于亲和纯化带有仅三个组氨酸残基的非常短标签的重组蛋白，适合于结晶。现在，实验结构是设计具有更高稳定性和亲和力的抗体的基础。 （c）2002爱思唯尔科学有限公司。

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