SARS-CoV-2 Spike Protein S2 Subunit: Recombinant Protein Expression Analysis, Purification, and Its Regulatory Effect...

Area of Science:

• Virology
• Molecular Biology
• Immunology

Background:

• The SARS-CoV-2 spike protein mediates viral entry via its S1 and S2 subunits.
• The S1 subunit binds ACE2, while the S2 subunit facilitates membrane fusion.
• The conserved S2 subunit is understudied despite its potential in host response modulation.

Purpose of the Study:

• To investigate the recombinant expression, purification, and functional impact of the SARS-CoV-2 S2 subunit.
• To explore the S2 subunit's role in modulating host cellular receptors.

Main Methods:

• Cloning the S2 coding region into a pGEX2TK plasmid for GST fusion protein production.
• Expressing and purifying the recombinant S2 protein in E. coli using glutathione affinity chromatography.
• Transfecting S2 protein into HEK293 and WI-38 cells to analyze IGF-1R expression.

Main Results:

• High-purity recombinant S2 protein was successfully expressed and purified.
• Transfection of S2 protein led to the downregulation of the insulin-like growth factor 1 receptor (IGF-1R).

Conclusions:

• The SARS-CoV-2 S2 subunit can modulate host cellular receptors, specifically downregulating IGF-1R.
• The S2 subunit's role in pathogenesis and therapeutic intervention warrants further investigation.