Complementing structural information of modular proteins with small angle neutron scattering and contrast variation

Eur Biophys J. 2008 Jun;37(5):603-11. doi: 10.1007/s00249-008-0278-z. Epub 2008 Feb 13.

Abstract

Many macromolecules in the cell function by forming multi-component assemblies. We have applied the technique of small angle neutron scattering to study a nucleic acid-protein complex and a multi-protein complex. The results illustrate the versatility and applicability of the method to study macromolecular assemblies. The neutron scattering experiments, complementing X-ray solution scattering data, reveal that the conserved catalytic domain of RNase E, an essential ribonuclease in Escherichia coli (E. coli), undergoes a marked conformational change upon binding a 5'monophosphate-RNA substrate analogue. This provides the first evidence in support of an allosteric mechanism that brings about RNA substrate cleavage. Neutron contrast variation of the multi-protein TIM10 complex, a mitochondrial chaperone assembly comprising the subunits Tim9 and Tim10, has been used to determine a low-resolution shape reconstruction of the complex, highlighting the integral subunit organization. It shows characteristic features involving protrusions that could be assigned to the six subunits forming the complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalytic Domain
  • Endoribonucleases / chemistry
  • Endoribonucleases / metabolism
  • Escherichia coli / enzymology
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism
  • Mitochondrial Precursor Protein Import Complex Proteins
  • Models, Molecular
  • Neutron Diffraction*
  • Nucleic Acids / chemistry*
  • Nucleic Acids / metabolism
  • Protein Binding
  • Proteins / chemistry*
  • Proteins / metabolism
  • RNA / metabolism
  • Saccharomyces cerevisiae Proteins / chemistry
  • Saccharomyces cerevisiae Proteins / metabolism
  • Scattering, Small Angle*
  • X-Ray Diffraction

Substances

  • Membrane Proteins
  • Mitochondrial Precursor Protein Import Complex Proteins
  • Nucleic Acids
  • Proteins
  • Saccharomyces cerevisiae Proteins
  • TIM10 protein, S cerevisiae
  • RNA
  • Endoribonucleases
  • ribonuclease E