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. 1993 Jan 1;289(Pt 1):87–91. doi: 10.1042/bj2890087

Purification of the pro-phenol oxidase enzyme from haemocytes of the cockroach Blaberus discoidalis.

H J Durrant 1, N A Ratcliffe 1, C R Hipkin 1, A Aspan 1, K Söderhäll 1
PMCID: PMC1132134  PMID: 8424776

Abstract

Pro-phenol oxidase was purified from the haemocytes of the cockroach Blaberus discoidalis by Blue Sepharose chromatography, hydrophobic-interaction chromatography on a Phenyl-Superose column and, finally, gel filtration on a Superose 6 column. Results suggest that the molecule exists as a polymer of identical 76 kDa monomeric units. The enzyme is a glycoprotein with pI of 5.2 and can be converted by trypsin into phenol oxidase.

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Selected References

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

  1. Asada N., Kitagawa O. Formation and the inhibition of reaction plug in mated Drosophila--study of a primitive defense reaction. Dev Comp Immunol. 1988 Summer;12(3):521–529. doi: 10.1016/0145-305x(88)90069-9. [DOI] [PubMed] [Google Scholar]
  2. Ashida M., Ishizaki Y., Iwahana H. Activation of pro-phenoloxidase by bacterial cell walls or beta-1,3-glucans in plasma of the silkworm, Bombyx mori. Biochem Biophys Res Commun. 1983 Jun 15;113(2):562–568. doi: 10.1016/0006-291x(83)91762-x. [DOI] [PubMed] [Google Scholar]
  3. Ashida M. Purification and characterization of pre-phenoloxidase from hemolymph of the silkworm Bombyx mori. Arch Biochem Biophys. 1971 Jun;144(2):749–762. doi: 10.1016/0003-9861(71)90383-3. [DOI] [PubMed] [Google Scholar]
  4. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  5. 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]
  6. Ferrini U., Mileo A. M., Hearing V. J. Microheterogeneity of melanosome-bound tyrosinase from the Harding-Passey murine melanoma. Int J Biochem. 1987;19(3):227–234. doi: 10.1016/0020-711x(87)90025-5. [DOI] [PubMed] [Google Scholar]
  7. Hiruma K., Riddiford L. M. Granular phenoloxidase involved in cuticular melanization in the tobacco hornworm: regulation of its synthesis in the epidermis by juvenile hormone. Dev Biol. 1988 Nov;130(1):87–97. doi: 10.1016/0012-1606(88)90416-2. [DOI] [PubMed] [Google Scholar]
  8. Imokawa G. Analysis of carbohydrate properties essential for melanogenesis in tyrosinases of cultured malignant melanoma cells by differential carbohydrate processing inhibition. J Invest Dermatol. 1990 Jul;95(1):39–49. doi: 10.1111/1523-1747.ep12873289. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Naqvi S. N., Karlson P. Purification of prophenoloxidase in the haemolymph of Calliphora vicina (R. & D.). Arch Int Physiol Biochim. 1979 Oct;87(4):687–695. doi: 10.3109/13813457909070529. [DOI] [PubMed] [Google Scholar]
  11. Nellaiappan K., Vinayagam A. A rapid method for detection of tyrosinase activity in electrophoresis. Stain Technol. 1986 Sep;61(5):269–272. doi: 10.3109/10520298609109952. [DOI] [PubMed] [Google Scholar]
  12. Ohkura T., Yamashita K., Mishima Y., Kobata A. Purification of hamster melanoma tyrosinases and structural studies of their asparagine-linked sugar chains. Arch Biochem Biophys. 1984 Nov 15;235(1):63–77. doi: 10.1016/0003-9861(84)90255-8. [DOI] [PubMed] [Google Scholar]
  13. Paskewitz S. M., Brown M. R., Collins F. H., Lea A. O. Ultrastructural localization of phenoloxidase in the midgut of refractory Anopheles gambiae and association of the enzyme with encapsulated Plasmodium cynomolgi. J Parasitol. 1989 Aug;75(4):594–600. [PubMed] [Google Scholar]
  14. Pye A. E. Microbial activation of prophenoloxidase from immune insect larvae. Nature. 1974 Oct 18;251(5476):610–613. doi: 10.1038/251610a0. [DOI] [PubMed] [Google Scholar]
  15. Ratcliffe N. A., Leonard C., Rowley A. F. Prophenoloxidase activation: nonself recognition and cell cooperation in insect immunity. Science. 1984 Nov 2;226(4674):557–559. doi: 10.1126/science.226.4674.557. [DOI] [PubMed] [Google Scholar]
  16. Rizki T. M., Rizki R. M., Bellotti R. A. Genetics of a Drosophila phenoloxidase. Mol Gen Genet. 1985;201(1):7–13. doi: 10.1007/BF00397978. [DOI] [PubMed] [Google Scholar]
  17. Saul S. J., Bin L., Sugumaran M. The majority of prophenoloxidase in the hemolymph of Manduca sexta is present in the plasma and not in the hemocytes. Dev Comp Immunol. 1987 Summer;11(3):479–485. doi: 10.1016/0145-305x(87)90037-1. [DOI] [PubMed] [Google Scholar]
  18. Saul S. J., Sugumaran M. Protease inhibitor controls prophenoloxidase activation in Manduca sexta. FEBS Lett. 1986 Nov 10;208(1):113–116. doi: 10.1016/0014-5793(86)81543-5. [DOI] [PubMed] [Google Scholar]
  19. Söderhäll K. Fungal cell wall beta-1,3-glucans induce clotting and phenoloxidase attachment to foreign surfaces of crayfish hemocyte lysate. Dev Comp Immunol. 1981 Fall;5(4):565–573. doi: 10.1016/s0145-305x(81)80031-6. [DOI] [PubMed] [Google Scholar]
  20. Söderhäll K. Prophenoloxidase activating system and melanization - a recognition mechanism of arthropods? A review. Dev Comp Immunol. 1982 Fall;6(4):601–611. [PubMed] [Google Scholar]
  21. Söderhäll K., Vey A., Ramstedt M. Hemocyte lysate enhancement of fungal spore encapsulation by crayfish hemocytes. Dev Comp Immunol. 1984 Winter;8(1):23–29. doi: 10.1016/0145-305x(84)90006-5. [DOI] [PubMed] [Google Scholar]
  22. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]

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