Skip to main content
PMC home page
Biochemical Journal logoLink to Biochemical Journal
. 1999 Jul 1;341(Pt 1):51–59.

Identification and expression of Pen c 2, a novel allergen from Penicillium citrinum.

L P Chow 1, N Y Su 1, C J Yu 1, B L Chiang 1, H D Shen 1
PMCID: PMC1220329  PMID: 10377244

Abstract

The mould genus, Penicillium, is known to be a significant source of environmental aero-allergens. One important allergen from Penicillium citrinum, Pen c 2, has been identified by means of two-dimensional immunoblotting using IgE-containing patients' sera. This novel allergen was cloned, sequenced and expressed in Escherichia coli. The cloned cDNA encodes a large 457-amino acid protein precursor containing a 16-amino acid signal peptide, a 120-amino acid propeptide and the 321-amino acid mature protein. Comparison of the Pen c 2 sequence with known protein sequences revealed shared high sequence similarities with two vacuolar serine proteases from Aspergillus niger and Saccharomyces cerevisiae. Asp-46, His-78 and Ser-244 were found to constitute the catalytic triad of the 39-kDa Pen c 2. The DNA coding for Pen c 2 was cloned into vector PQE-30 and expressed in E. coli as a His-tag fusion protein that bound serum IgE from Penicillium-allergic patients on immunoblots. Recombinant Pen c 2 could therefore be used effectively for diagnosis and also potentially for the treatment of mould-derived allergic disorders.

Full Text

The Full Text of this article is available as a PDF (551.7 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Belin L. G., Norman P. S. Diagnostic tests in the skin and serum of works sensitized to Bacillus subtilis enzymes. Clin Allergy. 1977 Jan;7(1):55–68. doi: 10.1111/j.1365-2222.1977.tb01425.x. [DOI] [PubMed] [Google Scholar]
  2. Betzel C., Pal G. P., Saenger W. Three-dimensional structure of proteinase K at 0.15-nm resolution. Eur J Biochem. 1988 Dec 1;178(1):155–171. doi: 10.1111/j.1432-1033.1988.tb14440.x. [DOI] [PubMed] [Google Scholar]
  3. Brouwer J. Cross-reactivity between Aspergillus fumigatus and Penicillium. Int Arch Allergy Immunol. 1996 Jun;110(2):166–173. doi: 10.1159/000237283. [DOI] [PubMed] [Google Scholar]
  4. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  5. Chow L.-P., Fukaya N., Miyatake N., Horimoto K., Sugiura Y., Tabuchi K., Ueno Y., Tsugita A. Resolution of Fusarium sporotrichioides Proteins by Two-Dimensional Polyacrylamide Gel Electrophoresis and Identification by Sequence Homology Comparison in Protein Data Base. J Biomed Sci. 1995 Oct;2(4):343–352. doi: 10.1007/BF02255221. [DOI] [PubMed] [Google Scholar]
  6. Flindt M. L. Pulmonary disease due to inhalation of derivatives of Bacillus subtilis containing proteolytic enzyme. Lancet. 1969 Jun 14;1(7607):1177–1181. doi: 10.1016/s0140-6736(69)92165-5. [DOI] [PubMed] [Google Scholar]
  7. Frederick G. D., Rombouts P., Buxton F. P. Cloning and characterisation of pepC, a gene encoding a serine protease from Aspergillus niger. Gene. 1993 Mar 15;125(1):57–64. doi: 10.1016/0378-1119(93)90745-o. [DOI] [PubMed] [Google Scholar]
  8. Hattori T., Ichihara S., Nakamura K. Processing of a plant vacuolar protein precursor in vitro. Eur J Biochem. 1987 Aug 3;166(3):533–538. doi: 10.1111/j.1432-1033.1987.tb13546.x. [DOI] [PubMed] [Google Scholar]
  9. Herbert C. A., King C. M., Ring P. C., Holgate S. T., Stewart G. A., Thompson P. J., Robinson C. Augmentation of permeability in the bronchial epithelium by the house dust mite allergen Der p1. Am J Respir Cell Mol Biol. 1995 Apr;12(4):369–378. doi: 10.1165/ajrcmb.12.4.7695916. [DOI] [PubMed] [Google Scholar]
  10. Horner W. E., Helbling A., Salvaggio J. E., Lehrer S. B. Fungal allergens. Clin Microbiol Rev. 1995 Apr;8(2):161–179. doi: 10.1128/cmr.8.2.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ikemura H., Takagi H., Inouye M. Requirement of pro-sequence for the production of active subtilisin E in Escherichia coli. J Biol Chem. 1987 Jun 5;262(16):7859–7864. [PubMed] [Google Scholar]
  12. Janknecht R., de Martynoff G., Lou J., Hipskind R. A., Nordheim A., Stunnenberg H. G. Rapid and efficient purification of native histidine-tagged protein expressed by recombinant vaccinia virus. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8972–8976. doi: 10.1073/pnas.88.20.8972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. King C., Brennan S., Thompson P. J., Stewart G. A. Dust mite proteolytic allergens induce cytokine release from cultured airway epithelium. J Immunol. 1998 Oct 1;161(7):3645–3651. [PubMed] [Google Scholar]
  14. King T. P., Hoffman D., Lowenstein H., Marsh D. G., Platts-Mills T. A., Thomas W. Allergen nomenclature. WHO/IUIS Allergen Nomenclature Subcommittee. Int Arch Allergy Immunol. 1994 Nov;105(3):224–233. doi: 10.1159/000236761. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Licorish K., Novey H. S., Kozak P., Fairshter R. D., Wilson A. F. Role of Alternaria and Penicillium spores in the pathogenesis of asthma. J Allergy Clin Immunol. 1985 Dec;76(6):819–825. doi: 10.1016/0091-6749(85)90755-9. [DOI] [PubMed] [Google Scholar]
  17. Moehle C. M., Tizard R., Lemmon S. K., Smart J., Jones E. W. Protease B of the lysosomelike vacuole of the yeast Saccharomyces cerevisiae is homologous to the subtilisin family of serine proteases. Mol Cell Biol. 1987 Dec;7(12):4390–4399. doi: 10.1128/mcb.7.12.4390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Sekimoto H., Seo M., Dohmae N., Takio K., Kamiya Y., Koshiba T. Cloning and molecular characterization of plant aldehyde oxidase. J Biol Chem. 1997 Jun 13;272(24):15280–15285. doi: 10.1074/jbc.272.24.15280. [DOI] [PubMed] [Google Scholar]
  19. Shen H. D., Lin W. L., Liaw S. F., Tam M. F., Han S. H. Characterization of the 33-kilodalton major allergen of Penicillium citrinum by using MoAbs and N-terminal amino acid sequencing. Clin Exp Allergy. 1997 Jan;27(1):79–86. [PubMed] [Google Scholar]
  20. Shen H. D., Lin W. L., Tam M. F., Wang S. R., Tsai J. J., Chou H., Han S. H. Alkaline serine proteinase: a major allergen of Aspergillus oryzae and its cross-reactivity with Penicillium citrinum. Int Arch Allergy Immunol. 1998 May;116(1):29–35. doi: 10.1159/000023921. [DOI] [PubMed] [Google Scholar]
  21. Shen H. D., Lin W. L., Tsai J. J., Liaw S. F., Han S. H. Allergenic components in three different species of Penicillium: crossreactivity among major allergens. Clin Exp Allergy. 1996 Apr;26(4):444–451. [PubMed] [Google Scholar]
  22. Smith W. A., Chua K. Y., Kuo M. C., Rogers B. L., Thomas W. R. Cloning and sequencing of the Dermatophagoides pteronyssinus group III allergen, Der p III. Clin Exp Allergy. 1994 Mar;24(3):220–228. doi: 10.1111/j.1365-2222.1994.tb00223.x. [DOI] [PubMed] [Google Scholar]
  23. Stevens T., Esmon B., Schekman R. Early stages in the yeast secretory pathway are required for transport of carboxypeptidase Y to the vacuole. Cell. 1982 Sep;30(2):439–448. doi: 10.1016/0092-8674(82)90241-0. [DOI] [PubMed] [Google Scholar]
  24. Stewart G. A., Boyd S. M., Bird C. H., Krska K. D., Kollinger M. R., Thompson P. J. Immunobiology of the serine protease allergens from house dust mites. Am J Ind Med. 1994 Jan;25(1):105–107. doi: 10.1002/ajim.4700250128. [DOI] [PubMed] [Google Scholar]
  25. Stewart G. A., Ward L. D., Simpson R. J., Thompson P. J. The group III allergen from the house dust mite Dermatophagoides pteronyssinus is a trypsin-like enzyme. Immunology. 1992 Jan;75(1):29–35. [PMC free article] [PubMed] [Google Scholar]
  26. Su N. Y., Yu C. J., Shen H. D., Pan F. M., Chow L. P. Pen c 1, a novel enzymic allergen protein from Penicillium citrinum. Purification, characterization, cloning and expression. Eur J Biochem. 1999 Apr;261(1):115–123. doi: 10.1046/j.1432-1327.1999.00242.x. [DOI] [PubMed] [Google Scholar]
  27. Verma J., Gangal S. V. Studies on Fusarium solani. Cross-reactivity among Fusarium species. Allergy. 1994 May;49(5):330–336. doi: 10.1111/j.1398-9995.1994.tb02277.x. [DOI] [PubMed] [Google Scholar]
  28. Zhu X. L., Ohta Y., Jordan F., Inouye M. Pro-sequence of subtilisin can guide the refolding of denatured subtilisin in an intermolecular process. Nature. 1989 Jun 8;339(6224):483–484. doi: 10.1038/339483a0. [DOI] [PubMed] [Google Scholar]
  29. von Heijne G. A new method for predicting signal sequence cleavage sites. Nucleic Acids Res. 1986 Jun 11;14(11):4683–4690. doi: 10.1093/nar/14.11.4683. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES