Conformational analysis of the GTP-binding protein MxA using limited proteolysis

FEBS Lett. 2002 Apr 10;516(1-3):129-32. doi: 10.1016/s0014-5793(02)02519-x.

Abstract

Guanosine triphosphate (GTP)-binding proteins are known to function as molecular switches that cycle between GTP-bound and guanosine diphosphate (GDP)-bound states. Switching is achieved by the fact that G-proteins in the GTP-bound conformation can interact with a certain set of effector molecules while they interact with a different set of partners in their GDP-bound conformation. The antiviral properties of the interferon-induced MxA protein are critically dependent on the ability of MxA to bind GTP. Using limited proteolysis we analyzed the conformations of the MxA protein under nucleotide-free, GDP-bound, and GTP-bound conditions. We find that whereas the conformations of nucleotide-free MxA and GDP-bound MxA are essentially similar, GTP-binding causes a dramatic change in the conformation of MxA.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiviral Agents / chemistry
  • Antiviral Agents / metabolism
  • Endopeptidase K
  • GTP-Binding Proteins / chemistry*
  • GTP-Binding Proteins / metabolism
  • Guanosine Diphosphate / metabolism
  • Guanosine Triphosphate / metabolism
  • In Vitro Techniques
  • Myxovirus Resistance Proteins
  • Papain
  • Protein Binding
  • Protein Conformation
  • Proteins / chemistry*
  • Proteins / metabolism
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / metabolism
  • Trypsin

Substances

  • Antiviral Agents
  • Myxovirus Resistance Proteins
  • Proteins
  • Recombinant Proteins
  • Guanosine Diphosphate
  • Guanosine Triphosphate
  • Trypsin
  • Endopeptidase K
  • Papain
  • GTP-Binding Proteins