Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1989 May 1;169(5):1757–1769. doi: 10.1084/jem.169.5.1757

Molecular cloning of the human eosinophil peroxidase. Evidence for the existence of a peroxidase multigene family

PMCID: PMC2189302  PMID: 2541222

Abstract

Human eosinophil peroxidase (EPO) was purified from eosinophil granules derived from the peripheral blood of patients with eosinophilia. The molecular mass of the H and L subunits was determined by gel filtration to be 57,000 and 11,000 daltons, respectively. The partial amino acid sequences of both subunits were used to construct oligonucleotides for the screening of several cDNA libraries, including one derived from human-induced umbilical cord mononuclear cells. A cDNA clone was isolated corresponding to EPO. The nucleotide sequence revealed an open reading frame of 2,106 bp, corresponding to a prosequence, L chain, and H chain, in this order. Comparison of the EPO nucleotide sequence with other peroxidases, such as myeloperoxidase, suggests the existence of a multigene family.

Full Text

The Full Text of this article is available as a PDF (1,012.3 KB).

Selected References

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

  1. Aviv H., Leder P. Purification of biologically active globin messenger RNA by chromatography on oligothymidylic acid-cellulose. Proc Natl Acad Sci U S A. 1972 Jun;69(6):1408–1412. doi: 10.1073/pnas.69.6.1408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benton W. D., Davis R. W. Screening lambdagt recombinant clones by hybridization to single plaques in situ. Science. 1977 Apr 8;196(4286):180–182. doi: 10.1126/science.322279. [DOI] [PubMed] [Google Scholar]
  3. Bolscher B. G., Plat H., Wever R. Some properties of human eosinophil peroxidase, a comparison with other peroxidases. Biochim Biophys Acta. 1984 Jan 31;784(2-3):177–186. doi: 10.1016/0167-4838(84)90125-0. [DOI] [PubMed] [Google Scholar]
  4. Carlson M. G., Peterson C. G., Venge P. Human eosinophil peroxidase: purification and characterization. J Immunol. 1985 Mar;134(3):1875–1879. [PubMed] [Google Scholar]
  5. Chirgwin J. M., Przybyla A. E., MacDonald R. J., Rutter W. J. Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry. 1979 Nov 27;18(24):5294–5299. doi: 10.1021/bi00591a005. [DOI] [PubMed] [Google Scholar]
  6. Fang G. H., Kenigsberg P., Axley M. J., Nuell M., Hager L. P. Cloning and sequencing of chloroperoxidase cDNA. Nucleic Acids Res. 1986 Oct 24;14(20):8061–8071. doi: 10.1093/nar/14.20.8061. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Fischkoff S. A., Pollak A., Gleich G. J., Testa J. R., Misawa S., Reber T. J. Eosinophilic differentiation of the human promyelocytic leukemia cell line, HL-60. J Exp Med. 1984 Jul 1;160(1):179–196. doi: 10.1084/jem.160.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Fish W. W., Mann K. G., Tanford C. The estimation of polypeptide chain molecular weights by gel filtration in 6 M guanidine hydrochloride. J Biol Chem. 1969 Sep 25;244(18):4989–4994. [PubMed] [Google Scholar]
  9. Gleich G. J., Adolphson C. R. The eosinophilic leukocyte: structure and function. Adv Immunol. 1986;39:177–253. doi: 10.1016/s0065-2776(08)60351-x. [DOI] [PubMed] [Google Scholar]
  10. Gleich G. J., Loegering D. A., Bell M. P., Checkel J. L., Ackerman S. J., McKean D. J. Biochemical and functional similarities between human eosinophil-derived neurotoxin and eosinophil cationic protein: homology with ribonuclease. Proc Natl Acad Sci U S A. 1986 May;83(10):3146–3150. doi: 10.1073/pnas.83.10.3146. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Goetzl E. J. The conversion of leukotriene C4 to isomers of leukotriene B4 by human eosinophil peroxidase. Biochem Biophys Res Commun. 1982 May 31;106(2):270–275. doi: 10.1016/0006-291x(82)91105-6. [DOI] [PubMed] [Google Scholar]
  12. Henderson W. R., Chi E. Y., Klebanoff S. J. Eosinophil peroxidase-induced mast cell secretion. J Exp Med. 1980 Aug 1;152(2):265–279. doi: 10.1084/jem.152.2.265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Jackson R. J., Hunt T. Preparation and use of nuclease-treated rabbit reticulocyte lysates for the translation of eukaryotic messenger RNA. Methods Enzymol. 1983;96:50–74. doi: 10.1016/s0076-6879(83)96008-1. [DOI] [PubMed] [Google Scholar]
  14. Johnson K. R., Nauseef W. M., Care A., Wheelock M. J., Shane S., Hudson S., Koeffler H. P., Selsted M., Miller C., Rovera G. Characterization of cDNA clones for human myeloperoxidase: predicted amino acid sequence and evidence for multiple mRNA species. Nucleic Acids Res. 1987 Mar 11;15(5):2013–2028. doi: 10.1093/nar/15.5.2013. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Jong E. C., Klebanoff S. J. Eosinophil-mediated mammalian tumor cell cytotoxicity: role of the peroxidase system. J Immunol. 1980 Apr;124(4):1949–1953. [PubMed] [Google Scholar]
  16. Jong E. C., Mahmoud A. A., Klebanoff S. J. Peroxidase-mediated toxicity to schistosomula of Schistosoma mansoni. J Immunol. 1981 Feb;126(2):468–471. [PubMed] [Google Scholar]
  17. Kimura S., Kotani T., McBride O. W., Umeki K., Hirai K., Nakayama T., Ohtaki S. Human thyroid peroxidase: complete cDNA and protein sequence, chromosome mapping, and identification of two alternately spliced mRNAs. Proc Natl Acad Sci U S A. 1987 Aug;84(16):5555–5559. doi: 10.1073/pnas.84.16.5555. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kraft R., Tardiff J., Krauter K. S., Leinwand L. A. Using mini-prep plasmid DNA for sequencing double stranded templates with Sequenase. Biotechniques. 1988 Jun;6(6):544-6, 549. [PubMed] [Google Scholar]
  19. Lagrimini L. M., Burkhart W., Moyer M., Rothstein S. Molecular cloning of complementary DNA encoding the lignin-forming peroxidase from tobacco: Molecular analysis and tissue-specific expression. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7542–7546. doi: 10.1073/pnas.84.21.7542. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  21. Magnusson R. P., Gestautas J., Taurog A., Rapoport B. Molecular cloning of the structural gene for porcine thyroid peroxidase. J Biol Chem. 1987 Oct 15;262(29):13885–13888. [PubMed] [Google Scholar]
  22. Messing J. New M13 vectors for cloning. Methods Enzymol. 1983;101:20–78. doi: 10.1016/0076-6879(83)01005-8. [DOI] [PubMed] [Google Scholar]
  23. Motojima S., Frigas E., Loegering D. A., Gleich G. J. Toxicity of eosinophil cationic proteins for guinea pig tracheal epithelium in vitro. Am Rev Respir Dis. 1989 Mar;139(3):801–805. doi: 10.1164/ajrccm/139.3.801. [DOI] [PubMed] [Google Scholar]
  24. Olsson I., Persson A. M., Strömberg K., Winqvist I., Tai P. C., Spry C. J. Purification of eosinophil peroxidase and studies of biosynthesis and processing in human marrow cells. Blood. 1985 Nov;66(5):1143–1148. [PubMed] [Google Scholar]
  25. Presentey B., Joshua H. Peroxidase and phospholipid deficiency in human eosinophilic granulocytes--a marker in population genetics. Experientia. 1982 May 15;38(5):628–629. doi: 10.1007/BF02327088. [DOI] [PubMed] [Google Scholar]
  26. Saito H., Hatake K., Dvorak A. M., Leiferman K. M., Donnenberg A. D., Arai N., Ishizaka K., Ishizaka T. Selective differentiation and proliferation of hematopoietic cells induced by recombinant human interleukins. Proc Natl Acad Sci U S A. 1988 Apr;85(7):2288–2292. doi: 10.1073/pnas.85.7.2288. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sukenaga Y., Ishida K., Takeda T., Takagi K. cDNA sequence coding for human glutathione peroxidase. Nucleic Acids Res. 1987 Sep 11;15(17):7178–7178. doi: 10.1093/nar/15.17.7178. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Tien M., Tu C. P. Cloning and sequencing of a cDNA for a ligninase from Phanerochaete chrysosporium. Nature. 1987 Apr 2;326(6112):520–523. doi: 10.1038/326520a0. [DOI] [PubMed] [Google Scholar]
  31. Wasmoen T. L., Bell M. P., Loegering D. A., Gleich G. J., Prendergast F. G., McKean D. J. Biochemical and amino acid sequence analysis of human eosinophil granule major basic protein. J Biol Chem. 1988 Sep 5;263(25):12559–12563. [PubMed] [Google Scholar]
  32. Wever R., Hamers M. N., Weening R. S., Roos D. Characterization of the peroxidase in human eosinophils. Eur J Biochem. 1980 Jul;108(2):491–495. doi: 10.1111/j.1432-1033.1980.tb04746.x. [DOI] [PubMed] [Google Scholar]
  33. Zucker-Franklin D., Grusky G. The identification of eosinophil colonies in soft-agar cultures by differential staining for peroxidase. J Histochem Cytochem. 1976 Dec;24(12):1270–1272. doi: 10.1177/24.12.63511. [DOI] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES