6X6P

Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis

Summary for 6X6P

• Entry DOI: 10.2210/pdb6x6p/pdb
• EMDB information: 22078
• Descriptor: Spike glycoprotein, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose, 2-acetamido-2-deoxy-beta-D-glucopyranose (3 entities in total)
• Functional Keywords: coronavirus, sars-cov-2, sars-cov, spike glycoprotein, fusion protein, viral protein, trimer
• Biological source: Severe acute respiratory syndrome coronavirus 2 (2019-nCoV)
• Total number of polymer chains: 3
• Total formula weight: 434865.89

Authors

Herrera, N.G.,Morano, N.C.,Celikgil, A.,Georgiev, G.I.,Malonis, R.,Lee, J.H.,Tong, K.,Vergnolle, O.,Massimi, A.,Yen, L.Y.,Noble, A.J.,Kopylov, M.,Bonanno, J.B.,Garrett-Thompson, S.C.,Hayes, D.B.,Brenowitz, M.,Garforth, S.J.,Eng, E.T.,Lai, J.R.,Almo, S.C. (deposition date: 2020-05-28, release date: 2020-06-10, Last modification date: 2024-10-23)

Primary citation

Herrera, N.G.,Morano, N.C.,Celikgil, A.,Georgiev, G.I.,Malonis, R.J.,Lee, J.H.,Tong, K.,Vergnolle, O.,Massimi, A.B.,Yen, L.Y.,Noble, A.J.,Kopylov, M.,Bonanno, J.B.,Garrett-Thomson, S.C.,Hayes, D.B.,Bortz, R.H.,Wirchnianski, A.S.,Florez, C.,Laudermilch, E.,Haslwanter, D.,Fels, J.M.,Dieterle, M.E.,Jangra, R.K.,Barnhill, J.,Mengotto, A.,Kimmel, D.,Daily, J.P.,Pirofski, L.A.,Chandran, K.,Brenowitz, M.,Garforth, S.J.,Eng, E.T.,Lai, J.R.,Almo, S.C.
Characterization of the SARS-CoV-2 S Protein: Biophysical, Biochemical, Structural, and Antigenic Analysis.
Biorxiv, 2020

PubMed Abstract: Coronavirus disease 2019 ( ) is a global health crisis caused by the novel severe acute respiratory syndrome coronavirus 2 ( ), and there is a critical need to produce large quantities of high-quality SARS-CoV-2 Spike ( ) 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 ( ) 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.

PubMed: 32587972
DOI: 10.1101/2020.06.14.150607

Experimental method

ELECTRON MICROSCOPY (3.22 Å)