Dimerization of recombinant tobacco mosaic virus movement protein

J Virol. 2004 Apr;78(7):3372-7. doi: 10.1128/jvi.78.5.3372-3377.2004.

Abstract

The p30 movement protein (MP) is essential for cell-to-cell spread of tobacco mosaic virus in planta. We used anion-exchange chromatography and preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) to obtain highly purified 30-kDa MP, which migrated as a single band in native PAGE. Analytical ultracentrifugation suggested that the protein was monodisperse and dimeric in the nonionic detergent n-octyl-beta-D-glucopyranoside. Circular dichroism (CD) spectroscopy showed that the detergent-solubilized protein contained significant alpha-helical secondary structure. Proteolysis of the C-tail generated a trypsin-resistant core that was a mixture of primarily monomers and some dimers. We propose that MP dimers are stabilized by electrostatic interactions in the C terminus as well as hydrophobic interactions between putative transmembrane alpha-helical coiled coils.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Circular Dichroism
  • Dimerization
  • Models, Molecular
  • Molecular Sequence Data
  • Plant Viral Movement Proteins
  • Protein Structure, Quaternary
  • Protein Structure, Secondary
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism
  • Solubility
  • Tobacco Mosaic Virus / chemistry*
  • Trypsin / metabolism
  • Ultracentrifugation
  • Viral Proteins / chemistry*
  • Viral Proteins / isolation & purification
  • Viral Proteins / metabolism*

Substances

  • Plant Viral Movement Proteins
  • Recombinant Proteins
  • Viral Proteins
  • Trypsin