5lem
From Proteopedia
(Difference between revisions)
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==Crystal structure of DARPin-DARPin rigid fusion, variant DD_Off7_11_3G124 in complex with Maltose-binding Protein and Green Fluorescent Protein== | ==Crystal structure of DARPin-DARPin rigid fusion, variant DD_Off7_11_3G124 in complex with Maltose-binding Protein and Green Fluorescent Protein== | ||
| - | <StructureSection load='5lem' size='340' side='right' caption='[[5lem]], [[Resolution|resolution]] 2.98Å' scene=''> | + | <StructureSection load='5lem' size='340' side='right'caption='[[5lem]], [[Resolution|resolution]] 2.98Å' scene=''> |
== Structural highlights == | == Structural highlights == | ||
| - | <table><tr><td colspan='2'>[[5lem]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Aeqvi Aeqvi], [http://en.wikipedia.org/wiki/ | + | <table><tr><td colspan='2'>[[5lem]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/Aeqvi Aeqvi], [http://en.wikipedia.org/wiki/Ecoli Ecoli] 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=5LEM OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5LEM FirstGlance]. <br> |
</td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr> | </td></tr><tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CRO:{2-[(1R,2R)-1-AMINO-2-HYDROXYPROPYL]-4-(4-HYDROXYBENZYLIDENE)-5-OXO-4,5-DIHYDRO-1H-IMIDAZOL-1-YL}ACETIC+ACID'>CRO</scene></td></tr> | ||
| - | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">malE, Z5632, ECs5017 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id= | + | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">malE, Z5632, ECs5017 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=83333 ECOLI]), GFP ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=6100 AEQVI])</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=5lem FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5lem OCA], [http://pdbe.org/5lem PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5lem RCSB], [http://www.ebi.ac.uk/pdbsum/5lem PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5lem ProSAT]</span></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=5lem FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5lem OCA], [http://pdbe.org/5lem PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5lem RCSB], [http://www.ebi.ac.uk/pdbsum/5lem PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=5lem ProSAT]</span></td></tr> | ||
</table> | </table> | ||
== Function == | == Function == | ||
[[http://www.uniprot.org/uniprot/GFP_AEQVI GFP_AEQVI]] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. [[http://www.uniprot.org/uniprot/MALE_ECO57 MALE_ECO57]] Involved in the high-affinity maltose membrane transport system MalEFGK. Initial receptor for the active transport of and chemotaxis toward maltooligosaccharides (By similarity). | [[http://www.uniprot.org/uniprot/GFP_AEQVI GFP_AEQVI]] Energy-transfer acceptor. Its role is to transduce the blue chemiluminescence of the protein aequorin into green fluorescent light by energy transfer. Fluoresces in vivo upon receiving energy from the Ca(2+)-activated photoprotein aequorin. [[http://www.uniprot.org/uniprot/MALE_ECO57 MALE_ECO57]] Involved in the high-affinity maltose membrane transport system MalEFGK. Initial receptor for the active transport of and chemotaxis toward maltooligosaccharides (By similarity). | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Multivalent binding proteins can gain biological activities beyond what is inherent in the individual binders, by bringing together different target molecules, restricting their conformational flexibility or changing their subcellular localization. In this study, we demonstrate a method to build up rigid multivalent and multispecific scaffolds by exploiting the modular nature of a repeat protein scaffold and avoiding flexible linkers. We use DARPins (Designed Ankyrin Repeat Proteins), synthetic binding proteins based on the Ankyrin-repeat protein scaffold, as binding units. Their ease of in vitro selection, high production yield and stability make them ideal specificity-conferring building blocks for the design of more complex constructs. C- and N-terminal DARPin capping repeats were re-designed to be joined by a shared helix in such a way that rigid connector modules are formed. This allows us to join two or more DARPins in predefined geometries without compromising their binding affinities and specificities. Nine connector modules with distinct geometries were designed; for eight of these we were able to confirm the structure by X-ray crystallography, while only one did not crystallize. The bispecific constructs were all able to bind both target proteins simultaneously. | ||
| + | |||
| + | Rigidly connected multispecific artificial binders with adjustable geometries.,Wu Y, Batyuk A, Honegger A, Brandl F, Mittl PRE, Pluckthun A Sci Rep. 2017 Sep 11;7(1):11217. doi: 10.1038/s41598-017-11472-x. PMID:28894181<ref>PMID:28894181</ref> | ||
| + | |||
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
| + | </div> | ||
| + | <div class="pdbe-citations 5lem" style="background-color:#fffaf0;"></div> | ||
| + | |||
| + | ==See Also== | ||
| + | *[[Green Fluorescent Protein|Green Fluorescent Protein]] | ||
| + | *[[Maltose-binding protein|Maltose-binding protein]] | ||
| + | == References == | ||
| + | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Aeqvi]] | [[Category: Aeqvi]] | ||
| - | [[Category: | + | [[Category: Ecoli]] |
| + | [[Category: Large Structures]] | ||
[[Category: Synthetic construct sequences]] | [[Category: Synthetic construct sequences]] | ||
[[Category: Batyuk, A]] | [[Category: Batyuk, A]] | ||
Revision as of 08:13, 20 March 2019
Crystal structure of DARPin-DARPin rigid fusion, variant DD_Off7_11_3G124 in complex with Maltose-binding Protein and Green Fluorescent Protein
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