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| | <StructureSection load='6i2g' size='340' side='right'caption='[[6i2g]], [[Resolution|resolution]] 1.50Å' scene=''> | | <StructureSection load='6i2g' size='340' side='right'caption='[[6i2g]], [[Resolution|resolution]] 1.50Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6i2g]] is a 2 chain structure with sequence from [http://en.wikipedia.org/wiki/Alpaca Alpaca]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6I2G OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6I2G FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6i2g]] is a 2 chain structure with sequence from [https://en.wikipedia.org/wiki/Vicugna_pacos Vicugna pacos] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6I2G OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6I2G FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><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]] 1.5Å</td></tr> |
| - | <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=LPD:L-PROLINAMIDE'>LPD</scene>, <scene name='pdbligand=N7P:1-ACETYL-L-PROLINE'>N7P</scene></td></tr> | + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=LPD:L-PROLINAMIDE'>LPD</scene>, <scene name='pdbligand=N7P:1-ACETYL-L-PROLINE'>N7P</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'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6i2g FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6i2g OCA], [http://pdbe.org/6i2g PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6i2g RCSB], [http://www.ebi.ac.uk/pdbsum/6i2g PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6i2g ProSAT]</span></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=6i2g FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6i2g OCA], [https://pdbe.org/6i2g PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6i2g RCSB], [https://www.ebi.ac.uk/pdbsum/6i2g PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6i2g ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: Alpaca]] | |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Martinez-Carranza, M]] | + | [[Category: Synthetic construct]] |
| - | [[Category: Stenmark, P]] | + | [[Category: Vicugna pacos]] |
| - | [[Category: Alfa]] | + | [[Category: Martinez-Carranza M]] |
| - | [[Category: Nanobody]] | + | [[Category: Stenmark P]] |
| - | [[Category: Peptide binding protein]]
| + | |
| - | [[Category: Tag]]
| + | |
| Structural highlights
Publication Abstract from PubMed
Specialized epitope tags are widely used for detecting, manipulating or purifying proteins, but often their versatility is limited. Here, we introduce the ALFA-tag, a rationally designed epitope tag that serves a remarkably broad spectrum of applications in life sciences while outperforming established tags like the HA-, FLAG(R)- or myc-tag. The ALFA-tag forms a small and stable alpha-helix that is functional irrespective of its position on the target protein in prokaryotic and eukaryotic hosts. We characterize a nanobody (NbALFA) binding ALFA-tagged proteins from native or fixed specimen with low picomolar affinity. It is ideally suited for super-resolution microscopy, immunoprecipitations and Western blotting, and also allows in vivo detection of proteins. We show the crystal structure of the complex that enabled us to design a nanobody mutant (NbALFA(PE)) that permits efficient one-step purifications of native ALFA-tagged proteins, complexes and even entire living cells using peptide elution under physiological conditions.
The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications.,Gotzke H, Kilisch M, Martinez-Carranza M, Sograte-Idrissi S, Rajavel A, Schlichthaerle T, Engels N, Jungmann R, Stenmark P, Opazo F, Frey S Nat Commun. 2019 Sep 27;10(1):4403. doi: 10.1038/s41467-019-12301-7. PMID:31562305[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Gotzke H, Kilisch M, Martinez-Carranza M, Sograte-Idrissi S, Rajavel A, Schlichthaerle T, Engels N, Jungmann R, Stenmark P, Opazo F, Frey S. The ALFA-tag is a highly versatile tool for nanobody-based bioscience applications. Nat Commun. 2019 Sep 27;10(1):4403. doi: 10.1038/s41467-019-12301-7. PMID:31562305 doi:http://dx.doi.org/10.1038/s41467-019-12301-7
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