title: Identification and Structural Characterization of an Intermediate in the Folding of the Measles Virus X Domain.
abstract: Although most proteins fold by populating intermediates, the transient nature of such states makes it difficult to characterize their structures. In this work we identified and characterized the structure of an intermediate of the X domain of phosphoprotein (P) of measles virus. We obtained this result by a combination of equilibrium and kinetic measurements and NMR chemical shifts used as structural restraints in replica-averaged metadynamics simulations. The structure of the intermediate was then validated by rationally designing four mutational variants predicted to affect the stability of this state. These results provide a detailed view of an intermediate state and illustrate the opportunities offered by a synergistic use of experimental and computational methods to describe non-native states at atomic resolution.
authors.affiliation: From the Istituto Pasteur-Fondazione Cenci Bolognetti and Istituto di Biologia e Patologia Molecolari del CNR, Dipartimento di Scienze Biochimiche "A. Rossi Fanelli," Sapienza Università di Roma, 00185 Rome, Italy.
authors.forename: Daniela
authors.initials: D
authors.lastname: Bonetti
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom, Department of Chemistry and Institute for Advanced Study, Technische Universität München, Lichtenbergstrasse 4, D-85747 Garching, Germany.
authors.forename: Carlo
authors.initials: C
authors.lastname: Camilloni
authors.affiliation: From the Istituto Pasteur-Fondazione Cenci Bolognetti and Istituto di Biologia e Patologia Molecolari del CNR, Dipartimento di Scienze Biochimiche "A. Rossi Fanelli," Sapienza Università di Roma, 00185 Rome, Italy.
authors.forename: Lorenzo
authors.initials: L
authors.lastname: Visconti
authors.affiliation: Aix-Marseille Université, Architecture et Fonction des Macromolécules Biologiques (AFMB), CNRS, UMR 7257, 13288 Marseille, France, and CNRS, AFMB UMR 7257, 13288 Marseille, France.
authors.forename: Sonia
authors.initials: S
authors.lastname: Longhi
authors.affiliation: From the Istituto Pasteur-Fondazione Cenci Bolognetti and Istituto di Biologia e Patologia Molecolari del CNR, Dipartimento di Scienze Biochimiche "A. Rossi Fanelli," Sapienza Università di Roma, 00185 Rome, Italy.
authors.forename: Maurizio
authors.initials: M
authors.lastname: Brunori
authors.affiliation: Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom.
authors.forename: Michele
authors.initials: M
authors.lastname: Vendruscolo
authors.affiliation: From the Istituto Pasteur-Fondazione Cenci Bolognetti and Istituto di Biologia e Patologia Molecolari del CNR, Dipartimento di Scienze Biochimiche "A. Rossi Fanelli," Sapienza Università di Roma, 00185 Rome, Italy, Department of Chemistry, University of Cambridge, Cambridge CB2 1EW, United Kingdom, stefano.gianni@uniroma1.it.
authors.forename: Stefano
authors.initials: S
authors.lastname: Gianni
mesh_terms: D008459:Measles virus / Q000737:chemistry*; D010750:Phosphoproteins / Q000737:chemistry*; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000737:chemistry*
chemical_list: C055859:P protein, Sendai virus; D010750:Phosphoproteins; D014764:Viral Proteins
keywords: fluorescence; kinetics; molecular dynamics; mutagenesis; nuclear magnetic resonance (NMR)