Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus
The nonstructural NS1 protein is an essential virulence factor of the human respiratory syncytial virus, with a predominant role in the inhibition of the host antiviral innate immune response. This inhibition is mediated by multiple protein-protein interactions and involves the formation of large ol...
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2015
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| Acceso en línea: | https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v54_n33_p5136_Pretel http://hdl.handle.net/20.500.12110/paper_00062960_v54_n33_p5136_Pretel |
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paper:paper_00062960_v54_n33_p5136_Pretel |
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dspace |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
Antibiotics Conformations Glycoproteins Immune system Life cycle Monomers Oligomers Polymerization Viruses Conformational heterogeneity Innate immune response Macromolecular crowding Protein concentrations Protein-protein interactions Respiratory syncytial virus Subcellular localizations Transition state ensembles Proteins amyloid monomer nonstructural protein 1 oligomer interferon protein binding virus protein Article beta sheet binding affinity cellular distribution controlled study half life time Human respiratory syncytial virus isomerization macromolecular crowding nonhuman observed rate constant oligomerization polymerization priority journal protein assembly protein binding protein conformation protein folding protein metabolism protein protein interaction protein quaternary structure protein secondary structure protein unfolding temperature chemistry enzyme specificity human Human respiratory syncytial virus kinetics metabolism physiology protein folding protein multimerization protein refolding solubility species difference Human respiratory syncytial virus Humans Interferons Kinetics Protein Binding Protein Folding Protein Multimerization Protein Refolding Protein Structure, Quaternary Protein Unfolding Respiratory Syncytial Virus, Human Solubility Species Specificity Substrate Specificity Temperature Viral Nonstructural Proteins |
| spellingShingle |
Antibiotics Conformations Glycoproteins Immune system Life cycle Monomers Oligomers Polymerization Viruses Conformational heterogeneity Innate immune response Macromolecular crowding Protein concentrations Protein-protein interactions Respiratory syncytial virus Subcellular localizations Transition state ensembles Proteins amyloid monomer nonstructural protein 1 oligomer interferon protein binding virus protein Article beta sheet binding affinity cellular distribution controlled study half life time Human respiratory syncytial virus isomerization macromolecular crowding nonhuman observed rate constant oligomerization polymerization priority journal protein assembly protein binding protein conformation protein folding protein metabolism protein protein interaction protein quaternary structure protein secondary structure protein unfolding temperature chemistry enzyme specificity human Human respiratory syncytial virus kinetics metabolism physiology protein folding protein multimerization protein refolding solubility species difference Human respiratory syncytial virus Humans Interferons Kinetics Protein Binding Protein Folding Protein Multimerization Protein Refolding Protein Structure, Quaternary Protein Unfolding Respiratory Syncytial Virus, Human Solubility Species Specificity Substrate Specificity Temperature Viral Nonstructural Proteins Chemes, Lucía Beatriz de Prat Gay, Gonzalo Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus |
| topic_facet |
Antibiotics Conformations Glycoproteins Immune system Life cycle Monomers Oligomers Polymerization Viruses Conformational heterogeneity Innate immune response Macromolecular crowding Protein concentrations Protein-protein interactions Respiratory syncytial virus Subcellular localizations Transition state ensembles Proteins amyloid monomer nonstructural protein 1 oligomer interferon protein binding virus protein Article beta sheet binding affinity cellular distribution controlled study half life time Human respiratory syncytial virus isomerization macromolecular crowding nonhuman observed rate constant oligomerization polymerization priority journal protein assembly protein binding protein conformation protein folding protein metabolism protein protein interaction protein quaternary structure protein secondary structure protein unfolding temperature chemistry enzyme specificity human Human respiratory syncytial virus kinetics metabolism physiology protein folding protein multimerization protein refolding solubility species difference Human respiratory syncytial virus Humans Interferons Kinetics Protein Binding Protein Folding Protein Multimerization Protein Refolding Protein Structure, Quaternary Protein Unfolding Respiratory Syncytial Virus, Human Solubility Species Specificity Substrate Specificity Temperature Viral Nonstructural Proteins |
| description |
The nonstructural NS1 protein is an essential virulence factor of the human respiratory syncytial virus, with a predominant role in the inhibition of the host antiviral innate immune response. This inhibition is mediated by multiple protein-protein interactions and involves the formation of large oligomeric complexes. There is neither a structure nor sequence or functional homologues of this protein, which points to a distinctive mechanism for blocking the interferon response among viruses. The NS1 native monomer follows a simple unfolding kinetics via a nativelike transition state ensemble, with a half-life of 45 min, in agreement with a highly stable core structure at equilibrium. Refolding is a complex process that involves several slowly interconverting species compatible with proline isomerization. However, an ultrafast folding event with a half-life of 0.2 ms is indicative of a highly folding compatible species within the unfolded state ensemble. On the other hand, the oligomeric assembly route from the native monomer, which does not involve unfolding, shows a monodisperse and irreversible end-point species triggered by a mild temperature change, with half-lives of 160 and 26 min at 37 and 47 °C, respectively, and at a low protein concentration (10 μM). A large secondary structure change into β-sheet structure and the formation of a dimeric nucleus precede polymerization by the sequential addition of monomers at the surprisingly low rate of one monomer every 34 s. The polymerization phase is followed by the binding to thioflavin-T indicative of amyloid-like, albeit soluble, repetitive β-sheet quaternary structure. The overall process is reversible only up until ∼8 min, a time window in which most of the secondary structure change takes place. NS1s multiple binding activities must be accommodated in a few binding interfaces at most, something to be considered remarkable given its small size (15 kDa). Thus, conformational heterogeneity, and in particular oligomer formation, may provide a means of expand its binding repertoire. These equilibria will be determined by variables such as macromolecular crowding, protein-protein interactions, expression levels, turnover, or specific subcellular localization. The irreversible and quasi-spontaneous nature of the oligomer assembly, together with the fact that NS1 is the most abundant viral protein in infected cells, makes its accumulation highly conceivable under conditions compatible with the cellular milieu. The implications of NS1 oligomers in the viral life cycle and the inhibition of host innate immune response remain to be determined. © 2015 American Chemical Society. |
| author |
Chemes, Lucía Beatriz de Prat Gay, Gonzalo |
| author_facet |
Chemes, Lucía Beatriz de Prat Gay, Gonzalo |
| author_sort |
Chemes, Lucía Beatriz |
| title |
Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus |
| title_short |
Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus |
| title_full |
Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus |
| title_fullStr |
Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus |
| title_full_unstemmed |
Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus |
| title_sort |
conformational heterogeneity determined by folding and oligomer assembly routes of the interferon response inhibitor ns1 protein, unique to human respiratory syncytial virus |
| publishDate |
2015 |
| url |
https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v54_n33_p5136_Pretel http://hdl.handle.net/20.500.12110/paper_00062960_v54_n33_p5136_Pretel |
| work_keys_str_mv |
AT chemesluciabeatriz conformationalheterogeneitydeterminedbyfoldingandoligomerassemblyroutesoftheinterferonresponseinhibitorns1proteinuniquetohumanrespiratorysyncytialvirus AT depratgaygonzalo conformationalheterogeneitydeterminedbyfoldingandoligomerassemblyroutesoftheinterferonresponseinhibitorns1proteinuniquetohumanrespiratorysyncytialvirus |
| _version_ |
1768544667131969536 |
| spelling |
paper:paper_00062960_v54_n33_p5136_Pretel2023-06-08T14:30:48Z Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus Chemes, Lucía Beatriz de Prat Gay, Gonzalo Antibiotics Conformations Glycoproteins Immune system Life cycle Monomers Oligomers Polymerization Viruses Conformational heterogeneity Innate immune response Macromolecular crowding Protein concentrations Protein-protein interactions Respiratory syncytial virus Subcellular localizations Transition state ensembles Proteins amyloid monomer nonstructural protein 1 oligomer interferon protein binding virus protein Article beta sheet binding affinity cellular distribution controlled study half life time Human respiratory syncytial virus isomerization macromolecular crowding nonhuman observed rate constant oligomerization polymerization priority journal protein assembly protein binding protein conformation protein folding protein metabolism protein protein interaction protein quaternary structure protein secondary structure protein unfolding temperature chemistry enzyme specificity human Human respiratory syncytial virus kinetics metabolism physiology protein folding protein multimerization protein refolding solubility species difference Human respiratory syncytial virus Humans Interferons Kinetics Protein Binding Protein Folding Protein Multimerization Protein Refolding Protein Structure, Quaternary Protein Unfolding Respiratory Syncytial Virus, Human Solubility Species Specificity Substrate Specificity Temperature Viral Nonstructural Proteins The nonstructural NS1 protein is an essential virulence factor of the human respiratory syncytial virus, with a predominant role in the inhibition of the host antiviral innate immune response. This inhibition is mediated by multiple protein-protein interactions and involves the formation of large oligomeric complexes. There is neither a structure nor sequence or functional homologues of this protein, which points to a distinctive mechanism for blocking the interferon response among viruses. The NS1 native monomer follows a simple unfolding kinetics via a nativelike transition state ensemble, with a half-life of 45 min, in agreement with a highly stable core structure at equilibrium. Refolding is a complex process that involves several slowly interconverting species compatible with proline isomerization. However, an ultrafast folding event with a half-life of 0.2 ms is indicative of a highly folding compatible species within the unfolded state ensemble. On the other hand, the oligomeric assembly route from the native monomer, which does not involve unfolding, shows a monodisperse and irreversible end-point species triggered by a mild temperature change, with half-lives of 160 and 26 min at 37 and 47 °C, respectively, and at a low protein concentration (10 μM). A large secondary structure change into β-sheet structure and the formation of a dimeric nucleus precede polymerization by the sequential addition of monomers at the surprisingly low rate of one monomer every 34 s. The polymerization phase is followed by the binding to thioflavin-T indicative of amyloid-like, albeit soluble, repetitive β-sheet quaternary structure. The overall process is reversible only up until ∼8 min, a time window in which most of the secondary structure change takes place. NS1s multiple binding activities must be accommodated in a few binding interfaces at most, something to be considered remarkable given its small size (15 kDa). Thus, conformational heterogeneity, and in particular oligomer formation, may provide a means of expand its binding repertoire. These equilibria will be determined by variables such as macromolecular crowding, protein-protein interactions, expression levels, turnover, or specific subcellular localization. The irreversible and quasi-spontaneous nature of the oligomer assembly, together with the fact that NS1 is the most abundant viral protein in infected cells, makes its accumulation highly conceivable under conditions compatible with the cellular milieu. The implications of NS1 oligomers in the viral life cycle and the inhibition of host innate immune response remain to be determined. © 2015 American Chemical Society. Fil:Chemes, L.B. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:De Prat-Gay, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2015 https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v54_n33_p5136_Pretel http://hdl.handle.net/20.500.12110/paper_00062960_v54_n33_p5136_Pretel |