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O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and <t>Ace2.</t> HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.
Human Ace2 Orf Cdna, supplied by Sino Biological, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/human+ace2+cdna+expression/Human+ACE2+%2F+Angiotensin-Converting+Enzyme+2+Gene+ORF+cDNA+clone+expression+plasmid%2C+C-HA+tag/pmc12403561-103-1-15
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O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and <t>Ace2.</t> HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.
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O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and <t>Ace2.</t> HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.
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O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and <t>Ace2.</t> HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.
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https://www.bioz.com/product/human+ace2+cdna+expression/Human+ACE2+%2F+Angiotensin-Converting+Enzyme+2+Gene+ORF+cDNA+clone+expression+plasmid%2C+C-His+tag/pm39856746-113-5-10
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O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and <t>Ace2.</t> HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.
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https://www.bioz.com/product/human+ace2+cdna+expression/Human+ACE2+%2F+Angiotensin-Converting+Enzyme+2+Gene+ORF+cDNA+clone+expression+plasmid%2C+C-His+tag/pm39856746-113-7-10
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O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and Ace2. HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.

Journal: Microbiology Spectrum

Article Title: O-GlcNAcylation at S659 enhances SARS-CoV-2 spike protein stability and pseudoparticle packaging efficiency

doi: 10.1128/spectrum.00527-25

Figure Lengend Snippet: O-GlcNAcylation of S at S659 regulates pseudoviral packaging of SARS-CoV-2. ( A ) S-S659A does not affect the binding between S and Ace2. HEK293T cells were transfected with the plasmids expressing HA-ACE2 and GFP-S-WT, GFP-S-659A, or empty vector. The cell lysates were subject to IP. ( B ) Quantitation of ( A ). ( C ) Diagram of four-plasmid system for SARS-CoV-2pp production. The four-plasmid system for preparing pseudovirus SARS-CoV-2pp includes Luc (pLenti6) encoding luciferase, HIV Gag/pol (pLP1), HIV Rev (pLP2), and GFP-S. ( D ) S659A mutation in SARS-CoV-2 S reduced the SARS-CoV2pp package efficiency. The luciferase activity of lysates from Huh7.5.1 cells infected with SARS-CoV-2pp generated from HEK293T cells based on either S or the S-S659A was assayed. ns, no significance; ****, P < 0.0001.

Article Snippet: The human ACE2 ORF cDNA clone expression plasmid with a C-HA tag was sourced from Sino Biological (HG10108-CY).

Techniques: Binding Assay, Transfection, Expressing, Plasmid Preparation, Quantitation Assay, Luciferase, Mutagenesis, Activity Assay, Infection, Generated

The model of O-GlcNAcylation of the SARS-CoV-2 S protein. The S protein is modified by O-GlcNAcylation, which is promoted by OGT and reduced by OGA. O-GlcNAcylation enhances the stability of the S protein, while non-GlcNAcylated S is targeted for ubiquitination and degradation. Thiamet G, an OGA inhibitor, elevates the levels of O-GlcNAcylation on the S protein. Proper glycosylation of the S protein is essential for the entry of SARS-CoV-2pp, which is mediated by the ACE2 receptor.

Journal: Microbiology Spectrum

Article Title: O-GlcNAcylation at S659 enhances SARS-CoV-2 spike protein stability and pseudoparticle packaging efficiency

doi: 10.1128/spectrum.00527-25

Figure Lengend Snippet: The model of O-GlcNAcylation of the SARS-CoV-2 S protein. The S protein is modified by O-GlcNAcylation, which is promoted by OGT and reduced by OGA. O-GlcNAcylation enhances the stability of the S protein, while non-GlcNAcylated S is targeted for ubiquitination and degradation. Thiamet G, an OGA inhibitor, elevates the levels of O-GlcNAcylation on the S protein. Proper glycosylation of the S protein is essential for the entry of SARS-CoV-2pp, which is mediated by the ACE2 receptor.

Article Snippet: The human ACE2 ORF cDNA clone expression plasmid with a C-HA tag was sourced from Sino Biological (HG10108-CY).

Techniques: Modification, Ubiquitin Proteomics, Glycoproteomics