Small-Angle Neutron Scattering for Structural Biology of Protein-RNA Complexes

Methods Enzymol. 2015:558:391-415. doi: 10.1016/bs.mie.2015.02.003. Epub 2015 Apr 1.

Abstract

This chapter deals with the applications of small-angle neutron scattering (SANS) for the structural study of protein-RNA complexes in solution. After a brief historical introduction, the basic theory and practical requirements (e.g., sample state) for SANS experiments will be treated. Next, model-free parameters, such as the molecular mass and the radius of gyration, which can be obtained without a priori structural information, will be introduced. A more detailed section on the specific properties of SANS (with respect to its sister technique, small-angle X-ray scattering), and their implications on possibilities and limits of model building and interpretation will be discussed with a focus on protein-RNA systems. A practical illustration of the information content of SANS data will be given by applying ab initio modeling to a tRNA-synthetase system of known high-resolution structure. Finally, two present state-of-the-art examples that combine SANS data with complementary structural biology techniques (NMR and crystallography) will be presented and possible future developments and applications will be discussed.

Keywords: Conformational change; Contrast variation; Deuteration; Modeling; Protein; RNA; Scattering length density; Shape; Small-angle neutron scattering.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acyl-tRNA Synthetases / chemistry*
  • Amino Acyl-tRNA Synthetases / metabolism
  • Animals
  • Binding Sites
  • Crystallography, X-Ray
  • DNA-Binding Proteins / chemistry*
  • DNA-Binding Proteins / metabolism
  • Drosophila Proteins / chemistry*
  • Drosophila Proteins / metabolism
  • Drosophila melanogaster
  • Humans
  • Magnetic Resonance Spectroscopy
  • Molecular Dynamics Simulation
  • Neutron Diffraction
  • Neutrons
  • Nuclear Proteins / chemistry*
  • Nuclear Proteins / metabolism
  • Nucleic Acid Conformation
  • Protein Binding
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • RNA / chemistry*
  • RNA / metabolism
  • RNA-Binding Proteins / chemistry*
  • RNA-Binding Proteins / metabolism
  • Ribonucleoproteins / chemistry*
  • Ribonucleoproteins / metabolism
  • Scattering, Small Angle
  • Solutions
  • Splicing Factor U2AF
  • Thermodynamics
  • Ubiquitin-Protein Ligases / chemistry*
  • Ubiquitin-Protein Ligases / metabolism

Substances

  • DNA-Binding Proteins
  • Drosophila Proteins
  • Nuclear Proteins
  • RNA-Binding Proteins
  • Ribonucleoproteins
  • Solutions
  • Splicing Factor U2AF
  • Sxl protein, Drosophila
  • U2AF2 protein, human
  • UNR protein, Drosophila
  • RNA
  • MSL2 protein, human
  • Ubiquitin-Protein Ligases
  • Amino Acyl-tRNA Synthetases