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Abstract:

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.

Registro:

Documento: Artículo
Título:Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus
Autor:Pretel, E.; Sánchez, I.E.; Fassolari, M.; Chemes, L.B.; De Prat-Gay, G.
Filiación:Protein Structure-Function and Engineering Laboratory, Fundación Instituto Leloir, IIBBA-CONICET, Av. Patricias Argentinas 435, Buenos Aires, 1405, Argentina
Protein Physiology Laboratory, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, IQUIBICEN-CONICET, Universidad de Buenos Aires, Buenos Aires, Argentina
Ciência Sem Fronteiras Senior Fellow, CNPq, Laboratório de Genômica Estrutural, Instituto de Biofísica Carlos Chagas Filho, Universidade Federal Do Rio de Janeiro, Rio de Janeiro, RJ, Brazil
Palabras clave: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
Año:2015
Volumen:54
Número:33
Página de inicio:5136
Página de fin:5146
DOI: http://dx.doi.org/10.1021/acs.biochem.5b00615
Título revista:Biochemistry
Título revista abreviado:Biochemistry
ISSN:00062960
CODEN:BICHA
CAS:amyloid, 11061-24-8; Interferons; Viral Nonstructural Proteins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_00062960_v54_n33_p5136_Pretel

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Citas:

---------- APA ----------
Pretel, E., Sánchez, I.E., Fassolari, M., Chemes, L.B. & De Prat-Gay, G. (2015) . Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus. Biochemistry, 54(33), 5136-5146.
http://dx.doi.org/10.1021/acs.biochem.5b00615
---------- CHICAGO ----------
Pretel, E., Sánchez, I.E., Fassolari, M., Chemes, L.B., De Prat-Gay, G. "Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus" . Biochemistry 54, no. 33 (2015) : 5136-5146.
http://dx.doi.org/10.1021/acs.biochem.5b00615
---------- MLA ----------
Pretel, E., Sánchez, I.E., Fassolari, M., Chemes, L.B., De Prat-Gay, G. "Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus" . Biochemistry, vol. 54, no. 33, 2015, pp. 5136-5146.
http://dx.doi.org/10.1021/acs.biochem.5b00615
---------- VANCOUVER ----------
Pretel, E., Sánchez, I.E., Fassolari, M., Chemes, L.B., De Prat-Gay, G. Conformational Heterogeneity Determined by Folding and Oligomer Assembly Routes of the Interferon Response Inhibitor NS1 Protein, Unique to Human Respiratory Syncytial Virus. Biochemistry. 2015;54(33):5136-5146.
http://dx.doi.org/10.1021/acs.biochem.5b00615