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. 1982 Dec 1;207(3):479–484. doi: 10.1042/bj2070479

Partial purification and characterization of an inhibitor from newborn-rat epidermis with activity against the proteinase of Schistosoma mansoni cercariae.

S Tzeng, J H McKerrow, K Jeong, K Fukuyama, W L Epstein
PMCID: PMC1153887  PMID: 7165704

Abstract

The penetration of cercariae through the skin initiates infection of the host with the human trematode parasite Schistosoma mansoni. Many larvae fail to migrate into the living epidermal cell layer. In order to determine if chemical as well as mechanical barriers to cercarial skin penetration exist, inhibitory activity of epidermal cell extracts against the proteinase obtained from cercarial secretions was assayed. An inhibitor was purified 50-fold by gel filtration on Sephadex G 75 and cation exchange chromatography at pH 5.8 and 4.9. The inhibitor has a relative molecular mass (Mr) of approx. 40 000-53 000. Oxidation of the inhibitor with N-chlorosuccinimide eliminated its inhibitory activity and thus indicated a critical methionine residue. The inhibitor was active against a wide spectrum of serine proteinases: porcine pancreatic elastase, human granulocyte elastase, bovine trypsin, and bovine alpha-chymotrypsin. However, no inhibition was detected against papain or clostridial collagenase. The inhibitor did not cross react with antiserum to human or rat serum alpha 1-proteinase inhibitor.

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Selected References

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

  1. Banda M. J., Werb Z. Mouse macrophage elastase. Purification and characterization as a metalloproteinase. Biochem J. 1981 Feb 1;193(2):589–605. doi: 10.1042/bj1930589. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bruce J. I., Pezzlo F., McCarty J. E., Yajima Y. Migration of Schistosoma mansoni through mouse tissue. Ultrastructure of host tissue and integument of migrating larva following cercarial penetration. Am J Trop Med Hyg. 1970 Nov;19(6):959–981. doi: 10.4269/ajtmh.1970.19.959. [DOI] [PubMed] [Google Scholar]
  3. Clegg J. A., Smithers S. R. Death of schistosome cercariae during penetration of the skin. II. Penetration of mammalian skin by Schistosoma mansoni. Parasitology. 1968 Feb;58(1):111–128. doi: 10.1017/s0031182000073479. [DOI] [PubMed] [Google Scholar]
  4. DAVIS B. J. DISC ELECTROPHORESIS. II. METHOD AND APPLICATION TO HUMAN SERUM PROTEINS. Ann N Y Acad Sci. 1964 Dec 28;121:404–427. doi: 10.1111/j.1749-6632.1964.tb14213.x. [DOI] [PubMed] [Google Scholar]
  5. Dresden M. H., Asch H. L. Proteolytic enzymes in extracts of Schistosoma mansoni cercariae. Biochim Biophys Acta. 1972 Dec 7;289(2):378–384. doi: 10.1016/0005-2744(72)90089-7. [DOI] [PubMed] [Google Scholar]
  6. Dresden M. H., Lewis J. C., Krisko I. Proteolytic action of Schistosoma mansoni cercarial proteases on keratin and basement membrane proteins. J Parasitol. 1977 Oct;63(5):941–943. [PubMed] [Google Scholar]
  7. Gazzinelli G., Ramalho-Pinto F. J., Pellegrino J. Purification and characterization of the proteolytic enzyme complex of cercarial extract. Comp Biochem Physiol. 1966 Aug;18(4):689–700. doi: 10.1016/0010-406x(66)90205-2. [DOI] [PubMed] [Google Scholar]
  8. Hibino T., Fukuyama K., Epstein W. L. Chemical characterization, synthesis and distribution of proteinase inhibitor in newborn rat epidermis. Biochim Biophys Acta. 1980 Oct 1;632(2):214–226. doi: 10.1016/0304-4165(80)90079-3. [DOI] [PubMed] [Google Scholar]
  9. Hibino T., Izaki S., Izaki M. Detection of serine proteinase inhibitors in human cornified cells. Biochem Biophys Res Commun. 1981 Aug 14;101(3):948–955. doi: 10.1016/0006-291x(81)91841-6. [DOI] [PubMed] [Google Scholar]
  10. Johnson D., Travis J. The oxidative inactivation of human alpha-1-proteinase inhibitor. Further evidence for methionine at the reactive center. J Biol Chem. 1979 May 25;254(10):4022–4026. [PubMed] [Google Scholar]
  11. Junnila S. K., Jansén C. T., Hopsu-Havu V. K. Elastase and trypsin inhibitors of human skin and serum. Partial purification and characterization. Acta Derm Venereol. 1971;51(4):251–256. [PubMed] [Google Scholar]
  12. Järvinen M. Purification and some characteristics of the human epidermal SH-protease inhibitor. J Invest Dermatol. 1978 Aug;71(2):114–118. doi: 10.1111/1523-1747.ep12546165. [DOI] [PubMed] [Google Scholar]
  13. Kominami E., Wakamatsu N., Katunuma N. Endogenous thiol protease inhibitor from rat liver. Biochem Biophys Res Commun. 1981 Mar 31;99(2):568–575. doi: 10.1016/0006-291x(81)91783-6. [DOI] [PubMed] [Google Scholar]
  14. LEWERT R. M., LEE C. L. Studies on the passage of helminth larvae through host tissues. I. Histochemical studies on the extracellular changes caused by penetrating larvae. II. Enzymatic activity of larvae in vitro and in vivo. J Infect Dis. 1954 Jul-Aug;95(1):13–51. doi: 10.1093/infdis/95.1.13. [DOI] [PubMed] [Google Scholar]
  15. Landsperger W. J., Stirewalt M. A., Dresden M. H. Purification and properties of a proteolytic enzyme from the cercariae of the human trematode parasite Schistosoma mansoni. Biochem J. 1982 Jan 1;201(1):137–144. doi: 10.1042/bj2010137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Lim H. K., Heyneman D. Intramolluscan inter-trematode antagonism: a review of factors influencing the host-parasite system and its possible role in biological control. Adv Parasitol. 1972;10:191–268. doi: 10.1016/s0065-308x(08)60175-x. [DOI] [PubMed] [Google Scholar]
  17. McKerrow J. H., Jeong K. H. Schistosoma mansoni: cercarial degradation of a radioactively labeled collagen gel. Exp Parasitol. 1982 Apr;53(2):249–254. doi: 10.1016/0014-4894(82)90066-2. [DOI] [PubMed] [Google Scholar]
  18. Roll D. E., Glew R. H. Isolation and characterization of rat alpha-1-antitrypsin. J Biol Chem. 1981 Aug 10;256(15):8190–8196. [PubMed] [Google Scholar]
  19. Smithers S. R., Terry R. J. The infection of laboratory hosts with cercariae of Schistosoma mansoni and the recovery of the adult worms. Parasitology. 1965 Nov;55(4):695–700. doi: 10.1017/s0031182000086248. [DOI] [PubMed] [Google Scholar]
  20. Stirewalt M. A., Austin B. E. Collection of secreted protease from the preacetabular glands of cercariae of Schistosoma mansoni. J Parasitol. 1973 Aug;59(4):741–743. [PubMed] [Google Scholar]
  21. Stirewalt M. A. Schistosoma mansoni: cercaria to schistosomule. Adv Parasitol. 1974;12:115–182. doi: 10.1016/s0065-308x(08)60388-7. [DOI] [PubMed] [Google Scholar]
  22. Stirewalt M. Quantitative collection and proteolytic activity of preacetabular gland enzyme (s) of cercariae of schistosoma mansoni. Am J Trop Med Hyg. 1978 May;27(3):548–553. doi: 10.4269/ajtmh.1978.27.548. [DOI] [PubMed] [Google Scholar]
  23. Takahara H., Nakayama H., Sinohara H. Purification and characterization of rat plasma alpha-1-antitrypsin. J Biochem. 1980 Aug;88(2):417–424. doi: 10.1093/oxfordjournals.jbchem.a132988. [DOI] [PubMed] [Google Scholar]
  24. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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