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The folding and aggregation properties of a single KH-domain protein: Ribosome binding factor A (RbfA) from Pseudomonas aeruginosa.
Santorelli, D; Rocchio, S; Fata, F; Silvestri, I; Angelucci, F; Imperi, F; Marasco, D; Diaferia, C; Gigli, L; Demitri, N; Federici, L; Di Matteo, A; Travaglini-Allocatelli, C.
Afiliación
  • Santorelli D; Department of Biochemical Sciences "A Rossi Fanelli" - Sapienza, University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy.
  • Rocchio S; Department of Biochemical Sciences "A Rossi Fanelli" - Sapienza, University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy; Institute of Molecular Biology and Pathology, National Research Council of Italy, c/o Department of Biochemical Sciences "A Rossi Fanelli" - Sapienza, University of Rome, P.le Al
  • Fata F; Department of Health, Life and Environmental Sciences, University of L'Aquila, P.le Salvatore Tommasi 1, 76100 L'Aquila, Italy.
  • Silvestri I; Department of Health, Life and Environmental Sciences, University of L'Aquila, P.le Salvatore Tommasi 1, 76100 L'Aquila, Italy.
  • Angelucci F; Department of Health, Life and Environmental Sciences, University of L'Aquila, P.le Salvatore Tommasi 1, 76100 L'Aquila, Italy.
  • Imperi F; Department of Science, Roma Tre University, Viale G. Marconi 446, 00146 Rome, Italy.
  • Marasco D; Department of Pharmacy, University of Naples "Federico II", Via Mezzocannone 16, 80134 Naples, Italy.
  • Diaferia C; Department of Pharmacy, University of Naples "Federico II", Via Mezzocannone 16, 80134 Naples, Italy.
  • Gigli L; Elettra - Sincrotrone Trieste, S.S. 14 Km 163.5, Area Science Park, 34149 Basovizza, Trieste, Italy.
  • Demitri N; Elettra - Sincrotrone Trieste, S.S. 14 Km 163.5, Area Science Park, 34149 Basovizza, Trieste, Italy.
  • Federici L; Department of Medical, Oral and Biotechnological Sciences and Center for Advanced Studies and Technology (CAST), University of Chieti "G. d'Annunzio", Via dei Vestini 31 - 66100, Chieti, Italy.
  • Di Matteo A; Institute of Molecular Biology and Pathology, National Research Council of Italy, c/o Department of Biochemical Sciences "A Rossi Fanelli" - Sapienza, University of Rome, P.le Aldo Moro 5, 00185, Rome, Italy. Electronic address: adele.dimatteo@cnr.it.
  • Travaglini-Allocatelli C; Department of Biochemical Sciences "A Rossi Fanelli" - Sapienza, University of Rome, P.le Aldo Moro 5, 00185 Rome, Italy. Electronic address: carlo.travaglini@uniroma1.it.
Biochim Biophys Acta Gen Subj ; 1865(2): 129780, 2021 02.
Article en En | MEDLINE | ID: mdl-33157160
BACKGROUND: Ribosome-binding factor A from the pathogenic bacterium Pseudomonas aeruginosa (PaRbfA) is a small ribosome assembly factor, composed by a single KH domain, involved in the maturation of the 30S subunit. These domains are characterized by the ability to bind RNA or ssDNA and are often located in proteins involved in a variety of cellular functions. However, although the ability of proteins to fold properly, to misfold or to aggregate is of paramount importance for their cellular functions, limited information is available on these dynamic properties in the case of KH domains. METHODS: PaRbfA thermodynamic stability and folding mechanism: Far-UV CD and fluorescence spectroscopy, stopped-flow kinetics and chevron plot analysis, site-directed mutagenesis. Fibrils characterization: FT-IR spectroscopy, Thioflavin T fluorescence, Transmission Electron Microscopy (TEM) and X-ray fibrils diffraction. RESULTS: Quantitative analysis of the (un)folding kinetics of PaRbfA show that, in vitro, the protein folds via a 3-states mechanism involving a transiently populated folding intermediate. We also provide experimental evidences that PaRbfA can form ordered fibrils endowed with cross-ß structure even in mild conditions. CONCLUSION: These results lead to the hypothesis that the folding intermediate of PaRbfA may expose (some of) the predicted amyloidogenic regions, which could act as aggregation nuclei in the fibrillogenesis. GENERAL SIGNIFICANCE: The methodological approach presented herein could be readily adapted to verify the ability of other KH domain proteins to form cross-ß structured fibrils and to transiently populate a folding intermediate.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pseudomonas aeruginosa Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Biochim Biophys Acta Gen Subj Año: 2021 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Países Bajos
Texto completo
Añadir a Mi BVS
Imprimir
XML
PubMed Links
Buscar en Google

Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Pseudomonas aeruginosa Tipo de estudio: Prognostic_studies Límite: Humans Idioma: En Revista: Biochim Biophys Acta Gen Subj Año: 2021 Tipo del documento: Article País de afiliación: Italia Pais de publicación: Países Bajos