Isolation and characterization of a proteinaceous inhibitor of microbial proteinases induced during the hypersensitive reaction of tobacco to tobacco mosaic virus

Mol Plant Microbe Interact. Sep-Oct 1990;3(5):327-33. doi: 10.1094/mpmi-3-327.

Abstract

A proteinase inhibitor is strongly induced in tobacco leaves reacting hypersensitively to tobacco mosaic virus. The tobacco inhibitor is highly active against four different serine endoproteinases of fungal and bacterial origin (EC 3.4.21.14) but inhibits poorly two serine endoproteinases of animal origin, trypsin (EC 3.4.21.4) and chymotrypsin (EC 3.4.21.1). The inhibitor has been purified to homogeneity by successive steps of conventional and high-performance liquid chromatography. When electrophoresed under denaturing conditions, it behaves as a small polypeptide with a molecular weight of about 6,000. From its amino acid composition and NH2-terminal amino acid sequence analysis, it appears that the inhibitor belongs to the potato inhibitor I family. A polyclonal antiserum was raised against the purified tobacco inhibitor and was used in immunoblotting experiments to follow inhibitor accumulation during the hypersensitive reaction of tobacco to tobacco mosaic virus. The inhibitor is highly efficient and might represent a potent fungicide and/or bactericide to be used in plant biotechnology.

MeSH terms

  • Amino Acid Sequence
  • Amino Acids / analysis
  • Immunoblotting
  • Molecular Sequence Data
  • Plant Diseases
  • Plant Proteins / chemistry
  • Plant Proteins / isolation & purification
  • Plant Proteins / metabolism*
  • Plants, Toxic*
  • Serine Proteinase Inhibitors / chemistry
  • Serine Proteinase Inhibitors / isolation & purification
  • Serine Proteinase Inhibitors / metabolism*
  • Substrate Specificity
  • Subtilisins / antagonists & inhibitors
  • Tobacco / immunology
  • Tobacco / microbiology*
  • Tobacco Mosaic Virus / physiology*

Substances

  • Amino Acids
  • PI-I protein, Nicotiana tabacum
  • Plant Proteins
  • Serine Proteinase Inhibitors
  • proteinase inhibitor I (plants)
  • Subtilisins