Artículo
Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants
Noguera, Martín Ezequiel
; Vazquez, Diego Sebastian; Ferrer Sueta, Gerardo; Agudelo Suarez, William Armando
; Howard, Eduardo Ignacio
; Rasia, Rodolfo Maximiliano
; Manta, Bruno; Cousido Siah, Alexandra; Mitschler, André; Podjarny, Alberto Daniel; Santos, Javier
Fecha de publicación:
02/2017
Editorial:
Nature Publishing Group
Revista:
Scientific Reports
ISSN:
2045-2322
Idioma:
Inglés
Tipo de recurso:
Artículo publicado
Clasificación temática:
Resumen
Thioredoxin is a ubiquitous small protein that catalyzes redox reactions of protein thiols. Additionally, thioredoxin from E. coli (EcTRX) is a widely-used model for structure-function studies. In a previous paper, we characterized several single-point mutants of the C-terminal helix (CTH) that alter global stability of EcTRX. However, spectroscopic signatures and enzymatic activity for some of these mutants were found essentially unaffected. A comprehensive structural characterization at the atomic level of these near-invariant mutants can provide detailed information about structural variability of EcTRX. We address this point through the determination of the crystal structures of four point-mutants, whose mutations occurs within or near the CTH, namely L94A, E101G, N106A and L107A. These structures are mostly unaffected compared with the wild-type variant. Notably, the E101G mutant presents a large region with two alternative traces for the backbone of the same chain. It represents a significant shift in backbone positions. Enzymatic activity measurements and conformational dynamics studies monitored by NMR and molecular dynamic simulations show that E101G mutation results in a small effect in the structural features of the protein. We hypothesize that these alternative conformations represent samples of the native-state ensemble of EcTRX, specifically the magnitude and location of conformational heterogeneity.
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Articulos(IBR)
Articulos de INST.DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Articulos de INST.DE BIOLOGIA MOLECULAR Y CELULAR DE ROSARIO
Articulos(IQUIFIB)
Articulos de INST.DE QUIMICA Y FISICO-QUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Articulos de INST.DE QUIMICA Y FISICO-QUIMICA BIOLOGICAS "PROF. ALEJANDRO C. PALADINI"
Citación
Noguera, Martín Ezequiel; Vazquez, Diego Sebastian; Ferrer Sueta, Gerardo; Agudelo Suarez, William Armando; Howard, Eduardo Ignacio; et al.; Structural variability of E. coli thioredoxin captured in the crystal structures of single-point mutants; Nature Publishing Group; Scientific Reports; 7; 2-2017; 1-12; 42343
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