Improved Accuracy from Joint X-ray and NMR Refinement of a Protein-RNA Complex Structure.

Integrated experimental approaches play an increasingly important role in structural biology, taking advantage of the complementary information provided by different techniques. In particular, the combination of NMR data with X-ray diffraction patterns may provide accurate and precise information ab...

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Publicado en:Journal of the American Chemical Society Vol. 138; no. 5; pp. 1601 - 1611
Autores principales: Carlon, Azzurra, Ravera, Enrico, Hennig, Janosch, Parigi, Giacomo, Sattler, Michael, Luchinat, Claudio
Formato: Artículo
Publicado: American Chemical Society 2/10/2016
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Acceso en línea:Ver este registro en EBSCOhost
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      dt: 2/10/2016
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      pub: American Chemical Society
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        10.1021/jacs.5b11598
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        atl: Improved Accuracy from Joint X-ray and NMR Refinement of a Protein-RNA Complex Structure.
      aug:
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          Carlon, Azzurra
          Ravera, Enrico
          Hennig, Janosch
          Parigi, Giacomo
          Sattler, Michael
          Luchinat, Claudio
        affil:
          Magnetic Resonance Center "CERM" and Department of Chemistry "Ugo Schiff", University of Florence and Magnetic Resonance Consortium (CIRMMP), Via L. Sacconi 6, 50019 Sesto Fiorentino, Firenze, Italy
          Center for Integrated Protein Science Munich (CIPSM) at Department Chemie, Technische Universität München, 85747 Garching, Germany
          Institute of Structural Biology, Helmholtz Zentrum München, 85764 Neuherberg, Germany
      su:
        RNA-protein interactions
        Nuclear magnetic resonance spectroscopy
        X-ray diffraction
        Molecular conformation
        Molecular structure
        Messenger RNA
      sug:
        subj:
          RNA-protein interactions
          Nuclear magnetic resonance spectroscopy
          X-ray diffraction
          Molecular conformation
          Molecular structure
          Messenger RNA
      ab: Integrated experimental approaches play an increasingly important role in structural biology, taking advantage of the complementary information provided by different techniques. In particular, the combination of NMR data with X-ray diffraction patterns may provide accurate and precise information about local conformations not available from average-resolution X-ray structures alone. Here, we refined the structure of a ternary protein-protein-RNA complex comprising three domains, Sxl and Unr, bound to a single-stranded region derived in the msl2 mRNA. The joint X-ray and NMR refinement reveals that--despite the poor quality of the fit found for the original structural model--the NMR data can be largely accommodated within the uncertainty in the atom positioning (structural noise) from the primary X-ray data and that the overall domain arrangements and binding interfaces are preserved on passing from the crystalline state to the solution. The refinement highlights local conformational differences, which provide additional information on specific features of the structure. For example, conformational dynamics and heterogeneity observed at the interface between the CSD1 and the Sxl protein components in the ternary complex are revealed by the combination of NMR and crystallographic data. The joint refinement protocol offers unique opportunities to detect structural differences arising from various experimental conditions and reveals static or dynamic differences in the conformation of the biomolecule between the solution and the crystals.
      pubtype: Academic Journal
      doctype: Article
      src: R
    language: English
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          year: 2016
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