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| | <StructureSection load='5xhg' size='340' side='right'caption='[[5xhg]], [[Resolution|resolution]] 1.76Å' scene=''> | | <StructureSection load='5xhg' size='340' side='right'caption='[[5xhg]], [[Resolution|resolution]] 1.76Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[5xhg]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Lk3_transgenic_mice Lk3 transgenic mice]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5XHG OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5XHG FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[5xhg]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Mus_musculus Mus musculus]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5XHG OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5XHG FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=OAZ:(2-azidophenyl)methyl+hydrogen+carbonate'>OAZ</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.76Å</td></tr> |
| - | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5xhg FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5xhg OCA], [http://pdbe.org/5xhg PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5xhg RCSB], [http://www.ebi.ac.uk/pdbsum/5xhg PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5xhg ProSAT]</span></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=EDO:1,2-ETHANEDIOL'>EDO</scene>, <scene name='pdbligand=OAZ:(2-azidophenyl)methyl+hydrogen+carbonate'>OAZ</scene>, <scene name='pdbligand=PEG:DI(HYDROXYETHYL)ETHER'>PEG</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> |
| | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5xhg FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5xhg OCA], [https://pdbe.org/5xhg PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5xhg RCSB], [https://www.ebi.ac.uk/pdbsum/5xhg PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5xhg ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Lk3 transgenic mice]] | + | [[Category: Mus musculus]] |
| - | [[Category: Kuratani, M]] | + | [[Category: Kuratani M]] |
| - | [[Category: Sakamoto, K]] | + | [[Category: Sakamoto K]] |
| - | [[Category: Yanagisawa, T]] | + | [[Category: Yanagisawa T]] |
| - | [[Category: Yokoyama, S]] | + | [[Category: Yokoyama S]] |
| - | [[Category: Fab]]
| + | |
| - | [[Category: Immune system]]
| + | |
| Structural highlights
Publication Abstract from PubMed
The site-specific chemical conjugation of proteins, following synthesis with an expanded genetic code, promises to advance antibody-based technologies, including antibody drug conjugation and the creation of bispecific Fab dimers. The incorporation of non-natural amino acids into antibodies not only guarantees site specificity but also allows the use of bio-orthogonal chemistry. However, the efficiency of amino acid incorporation fluctuates significantly among different sites, thereby hampering the identification of useful conjugation sites. In this study, we applied the codon reassignment technology to achieve the robust and efficient synthesis of chemically functionalized antibodies containing N(epsilon)-(o-azidobenzyloxycarbonyl)-l-lysine (o-Az-Z-Lys) at defined positions. This lysine derivative has a bio-orthogonally reactive group at the end of a long side chain, enabling identification of multiple new positions in Fab-constant domains, allowing chemical conjugation with high efficiency. An X-ray crystallographic study of a Fab variant with o-Az-Z-Lys revealed high-level exposure of the azido group to solvent, with six of the identified positions subsequently used to engineer "Variabodies", a novel antibody format allowing various connections between two Fab molecules. Our findings indicated that some of the created Variabodies exhibited agonistic activity in cultured cells as opposed to the antagonistic nature of antibodies. These results showed that our approach greatly enhanced the availability of antibodies for chemical conjugation and might aid in the development of new therapeutic antibodies.
Extensive Survey of Antibody Invariant Positions for Efficient Chemical Conjugation Using Expanded Genetic Codes.,Kato A, Kuratani M, Yanagisawa T, Ohtake K, Hayashi A, Amano Y, Kimura K, Yokoyama S, Sakamoto K, Shiraishi Y Bioconjug Chem. 2017 Aug 16;28(8):2099-2108. doi:, 10.1021/acs.bioconjchem.7b00265. Epub 2017 Jul 20. PMID:28727448[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Kato A, Kuratani M, Yanagisawa T, Ohtake K, Hayashi A, Amano Y, Kimura K, Yokoyama S, Sakamoto K, Shiraishi Y. Extensive Survey of Antibody Invariant Positions for Efficient Chemical Conjugation Using Expanded Genetic Codes. Bioconjug Chem. 2017 Aug 16;28(8):2099-2108. doi:, 10.1021/acs.bioconjchem.7b00265. Epub 2017 Jul 20. PMID:28727448 doi:http://dx.doi.org/10.1021/acs.bioconjchem.7b00265
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