1ktr
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(New page: 200px<br /> <applet load="1ktr" size="450" color="white" frame="true" align="right" spinBox="true" caption="1ktr, resolution 2.7Å" /> '''Crystal Structure of...)
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Revision as of 07:28, 18 November 2007
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Crystal Structure of the Anti-His Tag Antibody 3D5 Single-Chain Fragment (scFv) in Complex with a Oligohistidine peptide
Overview
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.
About this Structure
1KTR is a Single protein structure of sequence from Mus musculus. Full crystallographic information is available from OCA.
Reference
Crystal structure of the anti-His tag antibody 3D5 single-chain fragment complexed to its antigen., Kaufmann M, Lindner P, Honegger A, Blank K, Tschopp M, Capitani G, Pluckthun A, Grutter MG, J Mol Biol. 2002 Apr 19;318(1):135-47. PMID:12054774
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