Artículo

Vazquez, D.S.; Sánchez, I.E.; Garrote, A.; Sica, M.P.; Santos, J. "The E. Coli thioredoxin folding mechanism: The key role of the C-terminal helix" (2015) Biochimica et Biophysica Acta - Proteins and Proteomics. 1854(2):127-137
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Abstract:

In this work, the unfolding mechanism of oxidized Escherichia Coli thioredoxin (EcTRX) was investigated experimentally and computationally. We characterized seven point mutants distributed along the C-terminal α-helix (CTH) and the preceding loop. The mutations destabilized the protein against global unfolding while leaving the native structure unchanged. Global analysis of the unfolding kinetics of all variants revealed a linear unfolding route with a high-energy on-pathway intermediate state flanked by two transition state ensembles TSE1 and TSE2. The experiments show that CTH is mainly unfolded in TSE1 and the intermediate and becomes structured in TSE2. Structure-based molecular dynamics are in agreement with these experiments and provide protein-wide structural information on transient states. In our model, EcTRX folding starts with structure formation in the β-sheet, while the protein helices coalesce later. As a whole, our results indicate that the CTH is a critical module in the folding process, restraining a heterogeneous intermediate ensemble into a biologically active native state and providing the native protein with thermodynamic and kinetic stability. © 2014 Elsevier B.V. All rights reserved.

Registro:

Documento: Artículo
Título:The E. Coli thioredoxin folding mechanism: The key role of the C-terminal helix
Autor:Vazquez, D.S.; Sánchez, I.E.; Garrote, A.; Sica, M.P.; Santos, J.
Filiación:Instituto de Química y Fisicoquímica Biológicas Prof. Alejandro C. Paladini, Universidad de Buenos Aires, Junín 956, Buenos Aires, 1113AAD, Argentina
Laboratorio de Fisiología de Proteínas, Departamento de Química Biológica, Universidad de Buenos Aires, Buenos Aires, C1428EGA, Argentina
CONICET e Instituto de Energía y Desarrollo Sustentable (IEDS), Centro Atómico Bariloche, Comisión Nacional de Energía Atómica (CNEA), Av. E. Bustillo km 9.500, San Carlos de Bariloche, Río Negro, Argentina
Palabras clave:Folding kinetics; Intermediate state; Molecular dynamics; Protein stability; Transition state ensemble; thioredoxin; thioredoxin; alpha helix; Article; beta sheet; carboxy terminal sequence; crystallization; Escherichia coli; genetic heterogeneity; molecular dynamics; nonhuman; protein conformation; protein folding; protein stability; protein unfolding; thermodynamics; chemistry; Escherichia coli; genetics; kinetics; point mutation; protein conformation; protein secondary structure; protein unfolding; Escherichia coli; Escherichia coli; Kinetics; Molecular Dynamics Simulation; Point Mutation; Protein Conformation; Protein Folding; Protein Structure, Secondary; Protein Unfolding; Thermodynamics; Thioredoxins
Año:2015
Volumen:1854
Número:2
Página de inicio:127
Página de fin:137
DOI: http://dx.doi.org/10.1016/j.bbapap.2014.11.004
Título revista:Biochimica et Biophysica Acta - Proteins and Proteomics
Título revista abreviado:Biochim. Biophys. Acta Proteins Proteomics
ISSN:15709639
CODEN:BBAPB
CAS:thioredoxin, 52500-60-4; Thioredoxins
Registro:https://bibliotecadigital.exactas.uba.ar/collection/paper/document/paper_15709639_v1854_n2_p127_Vazquez

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Citas:

---------- APA ----------
Vazquez, D.S., Sánchez, I.E., Garrote, A., Sica, M.P. & Santos, J. (2015) . The E. Coli thioredoxin folding mechanism: The key role of the C-terminal helix. Biochimica et Biophysica Acta - Proteins and Proteomics, 1854(2), 127-137.
http://dx.doi.org/10.1016/j.bbapap.2014.11.004
---------- CHICAGO ----------
Vazquez, D.S., Sánchez, I.E., Garrote, A., Sica, M.P., Santos, J. "The E. Coli thioredoxin folding mechanism: The key role of the C-terminal helix" . Biochimica et Biophysica Acta - Proteins and Proteomics 1854, no. 2 (2015) : 127-137.
http://dx.doi.org/10.1016/j.bbapap.2014.11.004
---------- MLA ----------
Vazquez, D.S., Sánchez, I.E., Garrote, A., Sica, M.P., Santos, J. "The E. Coli thioredoxin folding mechanism: The key role of the C-terminal helix" . Biochimica et Biophysica Acta - Proteins and Proteomics, vol. 1854, no. 2, 2015, pp. 127-137.
http://dx.doi.org/10.1016/j.bbapap.2014.11.004
---------- VANCOUVER ----------
Vazquez, D.S., Sánchez, I.E., Garrote, A., Sica, M.P., Santos, J. The E. Coli thioredoxin folding mechanism: The key role of the C-terminal helix. Biochim. Biophys. Acta Proteins Proteomics. 2015;1854(2):127-137.
http://dx.doi.org/10.1016/j.bbapap.2014.11.004