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. 1993 Jun;102(2):639–644. doi: 10.1104/pp.102.2.639

Purification and characterization from tobacco (Nicotiana tabacum) leaves of six small, wound-inducible, proteinase isoinhibitors of the potato inhibitor II family.

G Pearce 1, S Johnson 1, C A Ryan 1
PMCID: PMC158823  PMID: 8108514

Abstract

Six small molecular mass, wound-inducible trypsin and chymotrypsin inhibitor proteins from tobacco (Nicotiana tabacum) leaves were isolated to homogeneity. The isoinhibitors, cumulatively called tobacco trypsin inhibitor (TTI), have molecular masses of approximately 5500 to 5800 D, calculated from gel filtration analysis and amino acid content. The amino acid sequence of the entire 53 residues of one isoinhibitor, TTI-1, and the sequence of 36 amino acid residues from the N terminus of a second isoinhibitor, TTI-5, were determined. The two isoinhibitors differ only at residue 11, which is threonine in TTI-1 and lysine in TTI-5. The isoinhibitors are members of the potato inhibitor II family and show considerable identity with the small molecular mass members of this family, which include the eggplant inhibitor, two small molecular mass trypsin and chymotrypsin inhibitors from potatoes, and an inhibitor from pistils of the ornamental plant Nicotiana alata. Antibodies produced against the isoinhibitors in rabbits were used in radial immunoassays to quantify both the systemic wound inducibility of TTI in tobacco leaves and its constitutive levels in flowers.

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Selected References

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  1. Atkinson A. H., Heath R. L., Simpson R. J., Clarke A. E., Anderson M. A. Proteinase inhibitors in Nicotiana alata stigmas are derived from a precursor protein which is processed into five homologous inhibitors. Plant Cell. 1993 Feb;5(2):203–213. doi: 10.1105/tpc.5.2.203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brown W. E., Ryan C. A. Isolation and characterization of a wound-induced trypsin inhibitor from alfalfa leaves. Biochemistry. 1984 Jul 17;23(15):3418–3422. doi: 10.1021/bi00310a006. [DOI] [PubMed] [Google Scholar]
  3. Bryant J., Green T. R., Gurusaddaiah T., Ryan C. A. Proteinase inhibitor II from potatoes: isolation and characterization of its protomer components. Biochemistry. 1976 Aug 10;15(16):3418–3424. doi: 10.1021/bi00661a004. [DOI] [PubMed] [Google Scholar]
  4. Graham J. S., Pearce G., Merryweather J., Titani K., Ericsson L. H., Ryan C. A. Wound-induced proteinase inhibitors from tomato leaves. II. The cDNA-deduced primary structure of pre-inhibitor II. J Biol Chem. 1985 Jun 10;260(11):6561–6564. [PubMed] [Google Scholar]
  5. Greenblatt H. M., Ryan C. A., James M. N. Structure of the complex of Streptomyces griseus proteinase B and polypeptide chymotrypsin inhibitor-1 from Russet Burbank potato tubers at 2.1 A resolution. J Mol Biol. 1989 Jan 5;205(1):201–228. doi: 10.1016/0022-2836(89)90376-8. [DOI] [PubMed] [Google Scholar]
  6. Hass G. M., Hermodson M. A., Ryan C. A., Gentry L. Primary structures of two low molecular weight proteinase inhibitors from potatoes. Biochemistry. 1982 Feb 16;21(4):752–756. doi: 10.1021/bi00533a027. [DOI] [PubMed] [Google Scholar]
  7. Melville J. C., Ryan C. A. Chymotrypsin inhibitor I from potatoes. Large scale preparation and characterization of its subunit components. J Biol Chem. 1972 Jun 10;247(11):3445–3453. [PubMed] [Google Scholar]
  8. Pearce G., Ryan C. A. A rapid, large-scale method for purification of the metallo-carboxypeptidase inhibitor from potato tubers. Anal Biochem. 1983 Apr 1;130(1):223–225. doi: 10.1016/0003-2697(83)90673-5. [DOI] [PubMed] [Google Scholar]
  9. Pearce G., Strydom D., Johnson S., Ryan C. A. A polypeptide from tomato leaves induces wound-inducible proteinase inhibitor proteins. Science. 1991 Aug 23;253(5022):895–897. doi: 10.1126/science.253.5022.895. [DOI] [PubMed] [Google Scholar]
  10. Pearce G., Sy L., Russell C., Ryan C. A., Hass G. M. Isolation and characterization from potato tubers of two polypeptide inhibitors of serine proteinases. Arch Biochem Biophys. 1982 Feb;213(2):456–462. doi: 10.1016/0003-9861(82)90571-9. [DOI] [PubMed] [Google Scholar]
  11. Plunkett G., Senear D. F., Zuroske G., Ryan C. A. Proteinase inhibitors I and II from leaves of wounded tomato plants: purification and properties. Arch Biochem Biophys. 1982 Feb;213(2):463–472. doi: 10.1016/0003-9861(82)90572-0. [DOI] [PubMed] [Google Scholar]
  12. Ryan C. A. The search for the proteinase inhibitor-inducing factor, PIIF. Plant Mol Biol. 1992 May;19(1):123–133. doi: 10.1007/BF00015610. [DOI] [PubMed] [Google Scholar]
  13. Swank R. T., Munkres K. D. Molecular weight analysis of oligopeptides by electrophoresis in polyacrylamide gel with sodium dodecyl sulfate. Anal Biochem. 1971 Feb;39(2):462–477. doi: 10.1016/0003-2697(71)90436-2. [DOI] [PubMed] [Google Scholar]
  14. Trautman R., Cowan K. M., Wagner G. G. Data processing for radial immunodiffusion. Immunochemistry. 1971 Oct;8(10):901–916. doi: 10.1016/0019-2791(71)90429-0. [DOI] [PubMed] [Google Scholar]
  15. Walker-Simmons M., Ryan C. A. Wound-induced Accumulation of Trypsin Inhibitor Activities in Plant Leaves: Survey of Several Plant Genera. Plant Physiol. 1977 Mar;59(3):437–439. doi: 10.1104/pp.59.3.437. [DOI] [PMC free article] [PubMed] [Google Scholar]

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