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| ==Single-Particle reconstruction of DARP14 - A designed protein scaffold displaying ~17kDa DARPin proteins - Scaffold== | | ==Single-Particle reconstruction of DARP14 - A designed protein scaffold displaying ~17kDa DARPin proteins - Scaffold== |
| <StructureSection load='6c9i' size='340' side='right' caption='[[6c9i]], [[Resolution|resolution]] 3.09Å' scene=''> | | <SX load='6c9i' size='340' side='right' viewer='molstar' caption='[[6c9i]], [[Resolution|resolution]] 3.09Å' scene=''> |
| == Structural highlights == | | == Structural highlights == |
| <table><tr><td colspan='2'>[[6c9i]] is a 24 chain structure with sequence from [http://en.wikipedia.org/wiki/Pseae Pseae] and [http://en.wikipedia.org/wiki/Pyrococcus_horikoshii Pyrococcus horikoshii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6C9I OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6C9I FirstGlance]. <br> | | <table><tr><td colspan='2'>[[6c9i]] is a 24 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudomonas_aeruginosa_PAO1 Pseudomonas aeruginosa PAO1] and [https://en.wikipedia.org/wiki/Pyrococcus_horikoshii_OT3 Pyrococcus horikoshii OT3]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6C9I OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6C9I FirstGlance]. <br> |
| </td></tr><tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">PH0671 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=70601 Pyrococcus horikoshii]), PA1966 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=208964 PSEAE])</td></tr> | | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 3.09Å</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=6c9i FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6c9i OCA], [http://pdbe.org/6c9i PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6c9i RCSB], [http://www.ebi.ac.uk/pdbsum/6c9i PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6c9i 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=6c9i FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6c9i OCA], [https://pdbe.org/6c9i PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6c9i RCSB], [https://www.ebi.ac.uk/pdbsum/6c9i PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6c9i ProSAT]</span></td></tr> |
| </table> | | </table> |
| <div style="background-color:#fffaf0;">
| | == Function == |
| == Publication Abstract from PubMed == | | [https://www.uniprot.org/uniprot/O58404_PYRHO O58404_PYRHO] |
| Current single-particle cryo-electron microscopy (cryo-EM) techniques can produce images of large protein assemblies and macromolecular complexes at atomic level detail without the need for crystal growth. However, proteins of smaller size, typical of those found throughout the cell, are not presently amenable to detailed structural elucidation by cryo-EM. Here we use protein design to create a modular, symmetrical scaffolding system to make protein molecules of typical size suitable for cryo-EM. Using a rigid continuous alpha helical linker, we connect a small 17-kDa protein (DARPin) to a protein subunit that was designed to self-assemble into a cage with cubic symmetry. We show that the resulting construct is amenable to structural analysis by single-particle cryo-EM, allowing us to identify and solve the structure of the attached small protein at near-atomic detail, ranging from 3.5- to 5-A resolution. The result demonstrates that proteins considerably smaller than the theoretical limit of 50 kDa for cryo-EM can be visualized clearly when arrayed in a rigid fashion on a symmetric designed protein scaffold. Furthermore, because the amino acid sequence of a DARPin can be chosen to confer tight binding to various other protein or nucleic acid molecules, the system provides a future route for imaging diverse macromolecules, potentially broadening the application of cryo-EM to proteins of typical size in the cell.
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| Near-atomic cryo-EM imaging of a small protein displayed on a designed scaffolding system.,Liu Y, Gonen S, Gonen T, Yeates TO Proc Natl Acad Sci U S A. 2018 Mar 5. pii: 1718825115. doi:, 10.1073/pnas.1718825115. PMID:29507202<ref>PMID:29507202</ref>
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| From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br>
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| </div>
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| <div class="pdbe-citations 6c9i" style="background-color:#fffaf0;"></div>
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| == References ==
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| <references/>
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| __TOC__ | | __TOC__ |
| </StructureSection> | | </SX> |
| [[Category: Pseae]] | | [[Category: Large Structures]] |
| [[Category: Pyrococcus horikoshii]] | | [[Category: Pseudomonas aeruginosa PAO1]] |
| [[Category: Gonen, S]] | | [[Category: Pyrococcus horikoshii OT3]] |
| [[Category: Gonen, T]] | | [[Category: Gonen S]] |
| [[Category: Liu, Y]] | | [[Category: Gonen T]] |
| [[Category: Yeates, T O]] | | [[Category: Liu Y]] |
| [[Category: Darpin]]
| | [[Category: Yeates TO]] |
| [[Category: De novo protein]]
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| [[Category: Designed complex]]
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| [[Category: Scaffold]]
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| [[Category: Single-particle cryo-em]]
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