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| | ==Crystal structure of a DARPin in complex with ORF49 from Lactococcal phage TP901-1== | | ==Crystal structure of a DARPin in complex with ORF49 from Lactococcal phage TP901-1== |
| - | <StructureSection load='3hg0' size='340' side='right' caption='[[3hg0]], [[Resolution|resolution]] 2.10Å' scene=''> | + | <StructureSection load='3hg0' size='340' side='right'caption='[[3hg0]], [[Resolution|resolution]] 2.10Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[3hg0]] is a 4 chain structure with sequence from [http://en.wikipedia.org/wiki/Bacteriophage_tp901-1 Bacteriophage tp901-1] and [http://en.wikipedia.org/wiki/Synthetic_construct_sequences Synthetic construct sequences]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3HG0 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=3HG0 FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[3hg0]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Bacteriophage_tp901-1 Bacteriophage tp901-1] and [https://en.wikipedia.org/wiki/Synthetic_construct_sequences Synthetic construct sequences]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3HG0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=3HG0 FirstGlance]. <br> |
| - | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[2bkk|2bkk]], [[1svx|1svx]], [[2v5q|2v5q]], [[2p2c|2p2c]], [[2j8s|2j8s]], [[2foc|2foc]]</td></tr> | + | </td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat"><div style='overflow: auto; max-height: 3em;'>[[2bkk|2bkk]], [[1svx|1svx]], [[2v5q|2v5q]], [[2p2c|2p2c]], [[2j8s|2j8s]], [[2foc|2foc]]</div></td></tr> |
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">bpp, bppL ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=35345 Bacteriophage TP901-1])</td></tr> | + | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">bpp, bppL ([https://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=35345 Bacteriophage TP901-1])</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=3hg0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3hg0 OCA], [http://pdbe.org/3hg0 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=3hg0 RCSB], [http://www.ebi.ac.uk/pdbsum/3hg0 PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=3hg0 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=3hg0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=3hg0 OCA], [https://pdbe.org/3hg0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=3hg0 RCSB], [https://www.ebi.ac.uk/pdbsum/3hg0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=3hg0 ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | == Evolutionary Conservation == | | == Evolutionary Conservation == |
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| | </StructureSection> | | </StructureSection> |
| | [[Category: Bacteriophage tp901-1]] | | [[Category: Bacteriophage tp901-1]] |
| | + | [[Category: Large Structures]] |
| | [[Category: Synthetic construct sequences]] | | [[Category: Synthetic construct sequences]] |
| | [[Category: Blangy, S]] | | [[Category: Blangy, S]] |
| Structural highlights
Evolutionary Conservation
Check, as determined by ConSurfDB. You may read the explanation of the method and the full data available from ConSurf.
Publication Abstract from PubMed
Combinatorial libraries of designed ankyrin repeat proteins (DARPins) have been proven to be a valuable source of specific binding proteins, as they can be expressed at very high levels and are very stable. We report here the selection of DARPins directed against a macromolecular multiprotein complex, the baseplate BppUxBppL complex of the lactococcal phage TP901-1. Using ribosome display, we selected several DARPins that bound specifically to the tip of the receptor-binding protein (RBP, the BppL trimer). The three selected DARPins display high specificity and affinity in the low nanomolar range and bind with a stoichiometry of one DARPin per BppL trimer. The crystal structure of a DARPin complexed with the RBP was solved at 2.1 A resolution. The DARPinxRBP interface is of the concave (DARPin)-convex (RBP) type, typical of other DARPin protein complexes and different from what is observed with a camelid VHH domain, which penetrates the phage p2 RBP inter-monomer interface. Finally, phage infection assays demonstrated that TP901-1 infection of Lactococcus lactis cells was inhibited by each of the three selected DARPins. This study provides proof of concept for the possible use of DARPins to circumvent viral infection. It also provides support for the use of DARPins in co-crystallization, due to their rigidity and their ability to provide multiple crystal contacts.
Crystal structure and function of a DARPin neutralizing inhibitor of lactococcal phage TP901-1: comparison of DARPin and camelid VHH binding mode.,Veesler D, Dreier B, Blangy S, Lichiere J, Tremblay D, Moineau S, Spinelli S, Tegoni M, Pluckthun A, Campanacci V, Cambillau C J Biol Chem. 2009 Oct 30;284(44):30718-26. Epub 2009 Sep 9. PMID:19740746[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Veesler D, Dreier B, Blangy S, Lichiere J, Tremblay D, Moineau S, Spinelli S, Tegoni M, Pluckthun A, Campanacci V, Cambillau C. Crystal structure and function of a DARPin neutralizing inhibitor of lactococcal phage TP901-1: comparison of DARPin and camelid VHH binding mode. J Biol Chem. 2009 Oct 30;284(44):30718-26. Epub 2009 Sep 9. PMID:19740746 doi:10.1074/jbc.M109.037812
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