Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin
Аутори
Simović, AnaBićanin, Maša
Radomirović, Mirjana Ž.
Aćimovič, Jelena
Mitić, Dragana
Stanić-Vučinić, Dragana
Ćirković-Veličković, Tanja
Конференцијски прилог (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
The emergence of the novel coronavirus SARS-CoV-2 has attracted the attention of the whole scientific community. However, as there are significant concerns regarding the effectiveness of vaccines and drugs against novel SARS-CoV-2 variants, naturally derived broad-spectrum of antivirals seems to be precious adjuvant agents to assist in combat against this disease. Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from Spirullina, with strong anti-oxidative action. The role of disulfide bonds and thiol-disulfide balance in RBD is considered to play a significant role in the binding of S protein to ACE2 receptor. In RBD, in contrast to C480–C488 disulfide, which is thermodynamically stable, C379–C432 and C391–C525 disulfides are in dynamic equilibrium with their thiol states and, thus these two pairs of disulfides are more sensitive to changes in redox poise. Our study aimed to investigate impact of PCB on disulfide balance of RBD ...by redox proteomics and to investigate structural changes in the protein exposed to PCB. The effect of PCB on RBD secondary structures was examined by far-UV CD spectroscopy after titration of RBD with increasing concentrations of PCB. The presence of PCB had a pronounced effect on the spectral shape. RBD is dominantly composed of random coils and β-sheets. In the presence of PCB a slight increase of α-helical and random coils content, while the content of β-sheets and β-turns is decreased. Mapping redox-active disulfides and reactive cysteines in recombinant SARS-CoV-2 RBD was done using redox proteomics on both recombinant RBD and PCB-exposed RBD. A mass shift caused by alkylation of free Cys residues was detected on three Cys residues demonstrating disulfides C379–C432 and 432-391 to be semi-stable in both RBD and PCB-exposed RBD. Our results demonstrate that RBD exposed to PCB undergo structural changes but does not change the redox state of its critical semi-stable disulfides.
Acknowledgment: The authors acknowledge support of the special Research Fund (BOF) of Ghent University (grant number 01N01718) and the Ministry of Science, Innovation and technological development of the Republic of Serbia (Contract number: 451-03-68/2023-14/200168).
Кључне речи:
Phycocyanobilin / Spike protein / ACE2 receptor / Redox proteomicsИзвор:
ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy, 2023, 63-63Издавач:
- Italian Proteomics Association
Финансирање / пројекти:
- Special Research Fund (BOF) of Ghent University (grant number 01N01718)
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200168 (Универзитет у Београду, Хемијски факултет) (RS-MESTD-inst-2020-200168)
Напомена:
- Book of Abstracts
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - CONF AU - Simović, Ana AU - Bićanin, Maša AU - Radomirović, Mirjana Ž. AU - Aćimovič, Jelena AU - Mitić, Dragana AU - Stanić-Vučinić, Dragana AU - Ćirković-Veličković, Tanja PY - 2023 UR - http://cherry.chem.bg.ac.rs/handle/123456789/6418 AB - The emergence of the novel coronavirus SARS-CoV-2 has attracted the attention of the whole scientific community. However, as there are significant concerns regarding the effectiveness of vaccines and drugs against novel SARS-CoV-2 variants, naturally derived broad-spectrum of antivirals seems to be precious adjuvant agents to assist in combat against this disease. Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from Spirullina, with strong anti-oxidative action. The role of disulfide bonds and thiol-disulfide balance in RBD is considered to play a significant role in the binding of S protein to ACE2 receptor. In RBD, in contrast to C480–C488 disulfide, which is thermodynamically stable, C379–C432 and C391–C525 disulfides are in dynamic equilibrium with their thiol states and, thus these two pairs of disulfides are more sensitive to changes in redox poise. Our study aimed to investigate impact of PCB on disulfide balance of RBD by redox proteomics and to investigate structural changes in the protein exposed to PCB. The effect of PCB on RBD secondary structures was examined by far-UV CD spectroscopy after titration of RBD with increasing concentrations of PCB. The presence of PCB had a pronounced effect on the spectral shape. RBD is dominantly composed of random coils and β-sheets. In the presence of PCB a slight increase of α-helical and random coils content, while the content of β-sheets and β-turns is decreased. Mapping redox-active disulfides and reactive cysteines in recombinant SARS-CoV-2 RBD was done using redox proteomics on both recombinant RBD and PCB-exposed RBD. A mass shift caused by alkylation of free Cys residues was detected on three Cys residues demonstrating disulfides C379–C432 and 432-391 to be semi-stable in both RBD and PCB-exposed RBD. Our results demonstrate that RBD exposed to PCB undergo structural changes but does not change the redox state of its critical semi-stable disulfides. Acknowledgment: The authors acknowledge support of the special Research Fund (BOF) of Ghent University (grant number 01N01718) and the Ministry of Science, Innovation and technological development of the Republic of Serbia (Contract number: 451-03-68/2023-14/200168). PB - Italian Proteomics Association C3 - ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy T1 - Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin SP - 63 EP - 63 UR - https://hdl.handle.net/21.15107/rcub_cherry_6418 ER -
@conference{ author = "Simović, Ana and Bićanin, Maša and Radomirović, Mirjana Ž. and Aćimovič, Jelena and Mitić, Dragana and Stanić-Vučinić, Dragana and Ćirković-Veličković, Tanja", year = "2023", abstract = "The emergence of the novel coronavirus SARS-CoV-2 has attracted the attention of the whole scientific community. However, as there are significant concerns regarding the effectiveness of vaccines and drugs against novel SARS-CoV-2 variants, naturally derived broad-spectrum of antivirals seems to be precious adjuvant agents to assist in combat against this disease. Phycocyanobilin (PCB) is an open-chain tetrapyrrole chromophore of phycocyanin (PC), chromoprotein derived from Spirullina, with strong anti-oxidative action. The role of disulfide bonds and thiol-disulfide balance in RBD is considered to play a significant role in the binding of S protein to ACE2 receptor. In RBD, in contrast to C480–C488 disulfide, which is thermodynamically stable, C379–C432 and C391–C525 disulfides are in dynamic equilibrium with their thiol states and, thus these two pairs of disulfides are more sensitive to changes in redox poise. Our study aimed to investigate impact of PCB on disulfide balance of RBD by redox proteomics and to investigate structural changes in the protein exposed to PCB. The effect of PCB on RBD secondary structures was examined by far-UV CD spectroscopy after titration of RBD with increasing concentrations of PCB. The presence of PCB had a pronounced effect on the spectral shape. RBD is dominantly composed of random coils and β-sheets. In the presence of PCB a slight increase of α-helical and random coils content, while the content of β-sheets and β-turns is decreased. Mapping redox-active disulfides and reactive cysteines in recombinant SARS-CoV-2 RBD was done using redox proteomics on both recombinant RBD and PCB-exposed RBD. A mass shift caused by alkylation of free Cys residues was detected on three Cys residues demonstrating disulfides C379–C432 and 432-391 to be semi-stable in both RBD and PCB-exposed RBD. Our results demonstrate that RBD exposed to PCB undergo structural changes but does not change the redox state of its critical semi-stable disulfides. Acknowledgment: The authors acknowledge support of the special Research Fund (BOF) of Ghent University (grant number 01N01718) and the Ministry of Science, Innovation and technological development of the Republic of Serbia (Contract number: 451-03-68/2023-14/200168).", publisher = "Italian Proteomics Association", journal = "ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy", title = "Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin", pages = "63-63", url = "https://hdl.handle.net/21.15107/rcub_cherry_6418" }
Simović, A., Bićanin, M., Radomirović, M. Ž., Aćimovič, J., Mitić, D., Stanić-Vučinić, D.,& Ćirković-Veličković, T.. (2023). Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy Italian Proteomics Association., 63-63. https://hdl.handle.net/21.15107/rcub_cherry_6418
Simović A, Bićanin M, Radomirović MŽ, Aćimovič J, Mitić D, Stanić-Vučinić D, Ćirković-Veličković T. Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin. in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy. 2023;:63-63. https://hdl.handle.net/21.15107/rcub_cherry_6418 .
Simović, Ana, Bićanin, Maša, Radomirović, Mirjana Ž., Aćimovič, Jelena, Mitić, Dragana, Stanić-Vučinić, Dragana, Ćirković-Veličković, Tanja, "Redox status of critical disulfides of SARS-CoV-2 receptor-binding-domain exposed to bioactive chomophore phycocyanobilin" in ItPA HPS and SePA XVII International Congress: Proteomics and Metabolomics towards Global Health, November 29th – December 1st, 2023, Roma, Italy (2023):63-63, https://hdl.handle.net/21.15107/rcub_cherry_6418 .