6x6p
From Proteopedia
(Difference between revisions)
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==Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis== | ==Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis== | ||
| - | <StructureSection load='6x6p' size='340' side='right'caption='[[6x6p]]' scene=''> | + | <StructureSection load='6x6p' size='340' side='right'caption='[[6x6p]], [[Resolution|resolution]] 3.22Å' scene=''> |
== Structural highlights == | == Structural highlights == | ||
| - | <table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6X6P OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=6X6P FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6x6p]] is a 3 chain structure with sequence from [http://en.wikipedia.org/wiki/2019-ncov 2019-ncov]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6X6P OCA]. For a <b>guided tour on the structure components</b> use [http://proteopedia.org/fgij/fg.htm?mol=6X6P FirstGlance]. <br> |
| - | </td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=6x6p FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6x6p OCA], [http://pdbe.org/6x6p PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6x6p RCSB], [http://www.ebi.ac.uk/pdbsum/6x6p PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6x6p ProSAT]</span></td></tr> | + | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NAG:N-ACETYL-D-GLUCOSAMINE'>NAG</scene></td></tr> |
| + | <tr id='gene'><td class="sblockLbl"><b>[[Gene|Gene:]]</b></td><td class="sblockDat">S, 2 ([http://www.ncbi.nlm.nih.gov/Taxonomy/Browser/wwwtax.cgi?mode=Info&srchmode=5&id=2697049 2019-nCoV])</td></tr> | ||
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://proteopedia.org/fgij/fg.htm?mol=6x6p FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6x6p OCA], [http://pdbe.org/6x6p PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6x6p RCSB], [http://www.ebi.ac.uk/pdbsum/6x6p PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6x6p ProSAT]</span></td></tr> | ||
</table> | </table> | ||
| + | == Function == | ||
| + | [[http://www.uniprot.org/uniprot/SPIKE_SARS2 SPIKE_SARS2]] attaches the virion to the cell membrane by interacting with host receptor, initiating the infection (By similarity). Binding to human ACE2 receptor and internalization of the virus into the endosomes of the host cell induces conformational changes in the Spike glycoprotein (PubMed:32142651, PubMed:32075877, PubMed:32155444). Uses also human TMPRSS2 for priming in human lung cells which is an essential step for viral entry (PubMed:32142651). Proteolysis by cathepsin CTSL may unmask the fusion peptide of S2 and activate membranes fusion within endosomes.[HAMAP-Rule:MF_04099]<ref>PMID:32075877</ref> <ref>PMID:32142651</ref> <ref>PMID:32155444</ref> mediates fusion of the virion and cellular membranes by acting as a class I viral fusion protein. Under the current model, the protein has at least three conformational states: pre-fusion native state, pre-hairpin intermediate state, and post-fusion hairpin state. During viral and target cell membrane fusion, the coiled coil regions (heptad repeats) assume a trimer-of-hairpins structure, positioning the fusion peptide in close proximity to the C-terminal region of the ectodomain. The formation of this structure appears to drive apposition and subsequent fusion of viral and target cell membranes.[HAMAP-Rule:MF_04099] Acts as a viral fusion peptide which is unmasked following S2 cleavage occurring upon virus endocytosis.[HAMAP-Rule:MF_04099] | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Coronavirus disease 2019 ( COVID-19 ) is a global health crisis caused by the novel severe acute respiratory syndrome coronavirus 2 ( SARS-CoV-2 ), and there is a critical need to produce large quantities of high-quality SARS-CoV-2 Spike ( S ) protein for use in both clinical and basic science settings. To address this need, we have evaluated the expression and purification of two previously reported S protein constructs in Expi293F () and ExpiCHO-S () cells, two different cell lines selected for increased expression of secreted glycoproteins. We show that ExpiCHO-S () cells produce enhanced yields of both SARS-CoV-2 S proteins. Biochemical, biophysical, and structural ( cryo-EM ) characterization of the SARS-CoV-2 S proteins produced in both cell lines demonstrate that the reported purification strategy yields high quality S protein (non-aggregated, uniform material with appropriate biochemical and biophysical properties). Importantly, we show that multiple preparations of these two recombinant S proteins from either cell line exhibit identical behavior in two different serology assays. We also evaluate the specificity of S protein-mediated host cell binding by examining interactions with proposed binding partners in the human secretome. In addition, the antigenicity of these proteins is demonstrated by standard ELISAs, and in a flexible protein microarray format. Collectively, we establish an array of metrics for ensuring the production of high-quality S protein to support clinical, biological, biochemical, structural and mechanistic studies to combat the global pandemic caused by SARS-CoV-2. | ||
| + | |||
| + | Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis.,Herrera NG, Morano NC, Celikgil A, Georgiev GI, Malonis RJ, Lee JH, Tong K, Vergnolle O, Massimi AB, Yen LY, Noble AJ, Kopylov M, Bonanno JB, Garrett-Thomson SC, Hayes DB, Bortz RH, Wirchnianski AS, Florez C, Laudermilch E, Haslwanter D, Fels JM, Dieterle ME, Jangra RK, Barnhill J, Mengotto A, Kimmel D, Daily JP, Pirofski LA, Chandran K, Brenowitz M, Garforth SJ, Eng ET, Lai JR, Almo SC bioRxiv. 2020 Jun 17. doi: 10.1101/2020.06.14.150607. PMID:32587972<ref>PMID:32587972</ref> | ||
| + | |||
| + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | ||
| + | </div> | ||
| + | <div class="pdbe-citations 6x6p" style="background-color:#fffaf0;"></div> | ||
| + | == References == | ||
| + | <references/> | ||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
| + | [[Category: 2019-ncov]] | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
| - | [[Category: Almo | + | [[Category: Almo, S C]] |
| - | [[Category: Bonanno | + | [[Category: Bonanno, J B]] |
| - | [[Category: Brenowitz M]] | + | [[Category: Brenowitz, M]] |
| - | [[Category: Celikgil A]] | + | [[Category: Celikgil, A]] |
| - | [[Category: Eng | + | [[Category: Eng, E T]] |
| - | [[Category: Garforth | + | [[Category: Garforth, S J]] |
| - | [[Category: Garrett-Thompson | + | [[Category: Garrett-Thompson, S C]] |
| - | [[Category: Georgiev | + | [[Category: Georgiev, G I]] |
| - | [[Category: Hayes | + | [[Category: Hayes, D B]] |
| - | [[Category: Herrera | + | [[Category: Herrera, N G]] |
| - | [[Category: Kopylov M]] | + | [[Category: Kopylov, M]] |
| - | [[Category: Lai | + | [[Category: Lai, J R]] |
| - | [[Category: Lee | + | [[Category: Lee, J H]] |
| - | [[Category: Malonis R]] | + | [[Category: Malonis, R]] |
| - | [[Category: Massimi A]] | + | [[Category: Massimi, A]] |
| - | [[Category: Morano | + | [[Category: Morano, N C]] |
| - | [[Category: Noble | + | [[Category: Noble, A J]] |
| - | [[Category: Tong K]] | + | [[Category: Tong, K]] |
| - | [[Category: Vergnolle O]] | + | [[Category: Vergnolle, O]] |
| - | [[Category: Yen | + | [[Category: Yen, L Y]] |
| + | [[Category: Coronavirus]] | ||
| + | [[Category: Fusion protein]] | ||
| + | [[Category: Sars-cov]] | ||
| + | [[Category: Sars-cov-2]] | ||
| + | [[Category: Spike glycoprotein]] | ||
| + | [[Category: Trimer]] | ||
| + | [[Category: Viral protein]] | ||
Revision as of 14:42, 8 July 2020
Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis
| |||||||||||
Categories: 2019-ncov | Large Structures | Almo, S C | Bonanno, J B | Brenowitz, M | Celikgil, A | Eng, E T | Garforth, S J | Garrett-Thompson, S C | Georgiev, G I | Hayes, D B | Herrera, N G | Kopylov, M | Lai, J R | Lee, J H | Malonis, R | Massimi, A | Morano, N C | Noble, A J | Tong, K | Vergnolle, O | Yen, L Y | Coronavirus | Fusion protein | Sars-cov | Sars-cov-2 | Spike glycoprotein | Trimer | Viral protein
