1887

Abstract

An elastase inhibitor from (AFUEI) was isolated, and its biochemical properties and primary structure examined. The inhibitor was purified by column chromatography using DE52 cellulose and Sephadex G-75, and was found to be homogeneous as indicated by a single band following discontinuous PAGE and SDS-PAGE. A molecular mass of 7525.1 Da was observed by matrix-assisted desorption/ionization time-of-flight mass spectroscopy. The elastolytic activity of elastases from , and human leukocytes was inhibited by AFUEI. However, the elastolytic activity of porcine pancreas elastase, elastase and elastase from snake venom was not affected by AFUEI. No inhibitory effect of DTT or 2-mercaptoethanol on the elastase inhibitory activity of AFUEI was observed. The amino acid sequence of AFUEI peptides derived from digests utilizing clostripain was determined by Edman sequencing. AFUEI was composed of 68 aa and had a calculated molecular mass of 7526.2 Da. The search for amino acid homology with other proteins demonstrated that aa 1–68 of AFUEI are 100 % identical to aa 20–87 of the hypothetical protein AFUA 3G14940 of .

Loading

Article metrics loading...

/content/journal/jmm/10.1099/jmm.0.47789-0
2008-07-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/jmm/57/7/803.html?itemId=/content/journal/jmm/10.1099/jmm.0.47789-0&mimeType=html&fmt=ahah

References

  1. Blanco J. L., Hontecillas R., Bouza E., Blanco I., Pelaez T., Munoz P., Molina J. P., Garcia M. E. 2002; Correlation between the elastase activity index and invasiveness of clinical isolates of Aspergillus fumigatus . J Clin Microbiol 40:1811–1813 [CrossRef]
    [Google Scholar]
  2. Frosco M., Chase T. Jr, Macmillan J. D. 1992a; Purification and properties of the elastase from Aspergillus fumigatus . Infect Immun 60:728–734
    [Google Scholar]
  3. Frosco M., Fahed C., Chase T. Jr, Macmillan J. D. 1992b; Inhibition of Aspergillus fumigatus elastase with monoclonal antibodies produced by using denatured elastase as an immunogen. Infect Immun 60:735–741
    [Google Scholar]
  4. Hasegawa Y., Nikai T., Yoshikawa Y., Sugihara H., Ogawa K., Takagi K. 1994; Elastase activity from Aspergillus species and inhibition by ulinastatin. Jap J Med Mycol 35:293–298 [CrossRef]
    [Google Scholar]
  5. Hasegawa Y., Nikai T., Yamashita R., Yoshikawa Y., Sugihara H., Ogawa K., Mizuno M. 1995; Isolation and characterization of elastolytic proteinase from Aspergillus fumigatus . Jap J Med Mycol 36:235–243 [CrossRef]
    [Google Scholar]
  6. Hasegawa Y., Nikai T., Okumura Y., Ogawa K., Sugihara H. 1997; Characterization of elastolytic proteinase from Aspergillus flavus : comparison of the elastolytic proteinase from Aspergillus fumigatus . Jap J Med Mycol 38:53–59 [CrossRef]
    [Google Scholar]
  7. Inoue H., Kimura T., Kitahara S., Makabe O., Takahashi K. 1991; The gene and deduced protein sequences of the zymogene of Aspergillus niger acid proteinase A. J Biol Chem 266:19484–19489
    [Google Scholar]
  8. Kawabata K., Suzuki M., Sugitani M., Imaki K., Toda M., Miyamoto T. 1991; ONO-5046, a novel inhibitor of human neutrophil elastase. Biochem Biophys Res Commun 177:814–820 [CrossRef]
    [Google Scholar]
  9. Kolattukudy P. E., Lee J. D., Rogers L. M., Zimmerman P., Ceselski S., Fox B., Stein B., Copelan E. A. 1993; Evidence for possible involvement of an elastolytic serine protease in aspergillosis. Infect Immun 61:2357–2368
    [Google Scholar]
  10. Komori Y., Nonogaki T., Nikai T. 2001; Hemorrhagic activity and muscle damaging effect of Pseudomonas aeruginosa metalloproteinase (elastase). Toxicon 39:1327–1332 [CrossRef]
    [Google Scholar]
  11. Kothary M. H., Chase T. Jr, Macmillan J. D. 1984; Correlation of elastase production by some strains of Aspergillus fumigatus with ability to cause pulmonary invasive aspergillosis in mice. Infect Immun 43:320–325
    [Google Scholar]
  12. Larcher G., Bouchara J. P., Annaix V., Symoens F., Chabasse D., Tronchin G. 1992; Purification and characterization of a fibrinogenolytic serine proteinase from Aspergillus fumigatus culture filtrate. FEBS Lett 308:65–69 [CrossRef]
    [Google Scholar]
  13. Maesaki S., Kohno S., Tanaka K., Miyazaki H., Mitsutake K., Miyazaki T., Tomono K., Kaku M., Koga H., Hara K. 1993; A case of pulmonary aspergilloma successfully treated with combination therapy of intracavitary injection of amphotericin B and intravenous administration of urinastatin. Nihon Kyobu Shikkan Gakkai Zasshi 31:1327–1331 (in Japanese
    [Google Scholar]
  14. Monod M., Togni G., Rahalison L., Frenk E. 1991; Isolation and characterization of an extracellular alkaline protease of Aspergillus fumigatus . J Med Microbiol 35:23–28 [CrossRef]
    [Google Scholar]
  15. Monod M., Paris S., Sangland D., Jaton-Ogay K., Bille J., Latge J. P. 1993; Isolation and characterization of secreted metalloprotease of Aspergillus fumigatus . Infect Immun 61:4099–4104
    [Google Scholar]
  16. Morris D. L. 1948; Quantitative determination of carbohydrates with Dreywood's anthrone reagent. Science 107:254–255 [CrossRef]
    [Google Scholar]
  17. Nierman W. C., Pain A., Anderson M. J., Wortman J. R., Kim H. S., Arroyo J., Berriman M., Abe K., Archer D. B. other authors 2005; Genomic sequence of the pathogenic and allergenic filamentous fungus Aspergillus fumigatus . Nature 438:1151–1156 [CrossRef]
    [Google Scholar]
  18. Ogawa K. 1997; Treatment of chronic pulmonary aspergillosis. Kekkaku 72:59–64 (in Japanese
    [Google Scholar]
  19. Ogawa K., Taki F., Takagi K., Furui H., Tsuda M., Honda K., Sasaki T., Sasamoto M., Miwa T. 1992; Study of elastase production by Aspergillus species and inhibition of their elastase activity by drugs. Kokyuu 11:880–886 (in Japanese
    [Google Scholar]
  20. Okumura Y., Ogawa K., Nikai T. 2004; Elastase and elastase inhibitor from Aspergillus fumigatus , Aspergillus flavus and Aspergillus niger . J Med Microbiol 53:351–354 [CrossRef]
    [Google Scholar]
  21. Okumura Y., Ogawa K., Uchiya K., Nikai T. 2006; Isolation and characterization of a novel elastase inhibitor, AFLEI from Aspergillus flavus . Jap J Med Mycol 47:219–224 [CrossRef]
    [Google Scholar]
  22. Okumura Y., Ogawa K., Nikai T. 2007a; Pathogenicity of elastase from Aspergillus flavus . J Res Inst Meijo Univ 6:145–152 (in Japanese
    [Google Scholar]
  23. Okumura Y., Ogawa K., Uchiya K., Nikai T. 2007b; Characterization and primary structure of elastase inhibitor, AFLEI, from Aspergillus flavus . Jap J Med Mycol 48:13–18 [CrossRef]
    [Google Scholar]
  24. Reichard U., Buttner S., Eiffert H., Staib F., Ruchel R. 1990; Purification and characterization of an extracellular serine proteinase from Aspergillus fumigatus and its detection in tissue. J Med Microbiol 33:243–251 [CrossRef]
    [Google Scholar]
  25. Rhodes J. C., Amlung T. W., Miller M. S. 1990; Isolation and characterization of an elastinolytic proteinase from Aspergillus flavus . Infect Immun 58:2529–2534
    [Google Scholar]
  26. Sugimura S., Yoshida K., Oba H., Hashiguchi K., Nakajima M., Moriya O., Okimoto N., Niki Y., Soejima R. 1994; Two cases of invasive pulmonary aspergillosis in non-immunocompromised hosts. Nihon Kyobu Shikkan Gakkai Zasshi 32:1032–1037 (in Japanese
    [Google Scholar]
  27. Takahashi K., Inoue H., Sakai K., Kohama T., Kitahara S., Takishima K., Tanji M., Athauda S., Takahashi T. other authors 1991; The primary structure of Aspergillus niger acid proteinase A. J Biol Chem 266:19480–19483
    [Google Scholar]
  28. Weber K., Osborn M. 1969; The reliability of molecular weight determination by dodecyl sulphate-polyacrylamide gel electrophoresis. J Biol Chem 244:4406–4412
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jmm/10.1099/jmm.0.47789-0
Loading
/content/journal/jmm/10.1099/jmm.0.47789-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error