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Cleavage of vesicular stomatitis virus matrix protein prevents self-association and leads to crystallization

M Gaudier1, Y Gaudin, M Knossow

  • 1Laboratoire d'Enzymologie et Biochimie Structurales, CNRS, 91198 Gif sur Yvette Cedex, France.

Virology
|October 17, 2001
PubMed

Insights

Vesicular stomatitis virus matrix protein (M) self-association, crucial for virus budding, was investigated. Proteolysis revealed a surface peptide drives this aggregation, enabling crystallization of the M protein core.

Area of Science:

  • Virology
  • Structural Biology
  • Protein Chemistry

Background:

  • The matrix protein (M) of vesicular stomatitis virus (VSV) mediates virion budding from host cells.
  • M protein self-association is implicated in virus assembly but hinders structural studies like crystallization.

Purpose of the Study:

  • To investigate the molecular basis of M protein self-association.
  • To obtain soluble, non-aggregating fragments of M protein for structural analysis.

Main Methods:

  • Limited proteolysis using thermolysin to digest the M protein.
  • Characterization of resulting protein fragments.
  • Crystallization of the thermolysin-resistant M protein core.

Main Results:

  • Two soluble, noncovalently associated fragments were isolated after thermolysin digestion.
  • These fragments did not self-associate or aggregate with intact M.
  • A surface-exposed peptide, disrupted by thermolysin, was identified as responsible for M self-association.
  • The remaining M protein core was crystallized and diffracted to 2-Å resolution.

Conclusions:

  • The M protein's self-association is mediated by a specific, surface-exposed peptide.
  • Disruption of this peptide via proteolysis yields soluble, non-aggregating fragments.
  • The thermolysin-resistant M protein core can be crystallized, paving the way for detailed structural studies.

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