Rapid measurement of residual dipolar couplings for fast fold elucidation of proteins
 

Rapid measurement of residual dipolar couplings for fast fold elucidation of proteins


Abstract It has been demonstrated that protein folds can be determined using appropriate computational protocols with NMR chemical shifts as the sole source of experimental restraints. While such approaches are very promising they still suffer from low convergence resulting in long computation times to achieve accurate results. Here we present a suite of time- and sensitivity optimized NMR experiments for rapid measurement of up to six RDCs per residue. Including such an RDC data set, measured in less than 24 h on a single aligned protein sample, greatly improves convergence of the Rosetta-NMR protocol, allowing for overnight fold calculation of small proteins. We demonstrate the performance of our fast fold calculation approach for ubiquitin as a test case, and for two RNA-binding domains of the plant protein HYL1. Structure calculations based on simulated RDC data highlight the importance of an accurate and precise set of several complementary RDCs as additional input restraints for high-quality de novo structure determination.

• Content Type Journal Article
• Category Article
• Pages 1-10
• DOI 10.1007/s10858-011-9567-4
• Authors
  • Rodolfo M. Rasia, Institut de Biologie Structurale, Jean-Pierre Ebel CNRS/CEA/UJF, 41 rue Jules Horowitz, 38027 Grenoble Cedex, France
  • Ewen Lescop, Institut de Biologie Structurale, Jean-Pierre Ebel CNRS/CEA/UJF, 41 rue Jules Horowitz, 38027 Grenoble Cedex, France
  • Javier F. Palatnik, Instituto de BiologÃ*a Molecular y Celular de Rosario, Facultad de Ciencias BioquÃ*micas y Farmacéuticas, Universidad Nacional de Rosario, Rosario, Argentina
  • Jérôme Boisbouvier, Institut de Biologie Structurale, Jean-Pierre Ebel CNRS/CEA/UJF, 41 rue Jules Horowitz, 38027 Grenoble Cedex, France
  • Bernhard Brutscher, Institut de Biologie Structurale, Jean-Pierre Ebel CNRS/CEA/UJF, 41 rue Jules Horowitz, 38027 Grenoble Cedex, France

• • Journal Journal of Biomolecular NMR
  • Online ISSN 1573-5001
  • Print ISSN 0925-2738

Source: Journal of Biomolecular NMR