Genomic and cDNA Sequence of Prophenoloxidases From Drosophila Melanogaster

  • Michael R. Chase
  • Manickam Sugumaran
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 484)


Phenoloxidase (PO) is an important enzyme associated with the biochemistry of three physiologically important processes in insects. The first process, viz., cuticular sclerotization, ensures the protection of the soft bodied insects from their predators and preys as well as from desiccation by hardening the cuticle (Andersen et al.,1996; Sugumaran 1991; 1998). During sclerotization, PO generated quinones serve both as sclerotizing agents and as substrates for quinone isomerases that produce quinone methides (Sugumaran, 1998; Saul & Sugumaran 1988, 1989 a,b,1990, Ricketts & Sugumaran 1994). The reactions of quinones and quinone methides with cuticular structural components viz., chitin and proteins, lead to the hardening of cuticle (Andersen et al., 1996; Sugumaran 1991, 1998). In the second process, organisms establishing successful entry into the insect hemocel face among other host defense reactions, the dreaded action of PO, which causes deposition of melanin pigment around the intruder (Ashida & Brey 1995; Gillespie et al., 1997>; Nappi & Sugumaran 1993; Soderhall, et al., 1990; Sugumaran, 1996; Sugumaran & Kanost, 1993). While the intruder may still live in side the melanotic encapsule, its ability to multiply and damage the host is dramatically hindered by the melanotic capsule. Finally during wounding, loss of insect hemolymph is arrested by the rapid deposition melanin at the wounding site (Lai-Fook, 1966; Sugumaran, 1996). Apart from sealing the wound, PO action may also provide cytotoxic quinones that could harm the opportunistically invading microorganism (Nappi & Sugumaran 1993; >Sugumaran 1996).


Malaria Vector Phenol Oxidase Intron Position Quinone Methides Insect Hemolymph 
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  1. Andersen SO, Peter MG, Roepstorff P. Cuticular sclerotization in insects. Comp Biochem. Physiol. 1996; 113B: 689–705.Google Scholar
  2. Andersson K, Sun SC, Boman HG, Steiner H. Purification of the prophenoloxidase from Hyalophora cecropia and four proteins involved in its activation. Insect Biochem. 1989; 19: 629–637.CrossRefGoogle Scholar
  3. Ashida M. Purification and characterization of prephenoloxidase from hemolymph of the silkworm, Bombyx mori. Arch. Biochem. Biophys. 1971; 144: 749–762.PubMedCrossRefGoogle Scholar
  4. Ashida M, Brey P. Role of the integument in insect defense: Prophenoloxidase cascade in the cuticular matrix. Proc. Natl. Acad. Sci. USA. 1995; 92: 10698–10702.PubMedCrossRefGoogle Scholar
  5. Ashida M, Yamazaki HI. “Biochemistry of the phenoloxidase system in insects: With special reference to its activation”. In Molting and metamorphosis, Ohnishi E, Ishizaki H. eds. Tokyo. Jpn. Sci. Soc. Press. 1990. pp. 239–265.Google Scholar
  6. Aso Y, Kramer ICJ, Hopkins TL, Lookhart GL. Characterization of hemolymph protyrosinase and a cuticular activator from Manduca sexta (L). Insect Biochem. 1985; 15: 9–17.CrossRefGoogle Scholar
  7. Burge CB. “Modeling dependencies in pre-mRNA splicing signals”. In Computational Methods in Molecular Biology, Salzberg, S., Semis, D. and Kasif, S. eds. Amsterdam, Elsevier Science, 1998. pp. 127–163.Google Scholar
  8. Burge C, Karlin S. Prediction of complete gene structures in human genomic DNA. J. Mol. Biol. 1997; 268: 78–94.PubMedCrossRefGoogle Scholar
  9. Chase MR, Raina K, Bruno J, Sugumaran M. Purification, characterization and molecular cloning of prophenoloxidases from Sarcophaga bullata. Insect Biochem. Mol. Biol. (in press, 2000)Google Scholar
  10. Cherqui A, Duvic B, Brehelin M. Purification and characterization of prophenoloxidase from the hemolymph of Locusta migratoria. Arch. Insect Biochem. Physiol. 1996; 32: 225–235.CrossRefGoogle Scholar
  11. Cho WL, Liu HS, Lee CH, Kuo CC, Chang TY, Liu CT, Chen CC. Molecular cloning, characterization and tissue expression of prophenoloxidase eDNA from the mosquito Armigeres subalbatus inoculated with Dirofilaria immitis microfilariae. Insect Mol. Biol. 1998; 7: 31–40.PubMedCrossRefGoogle Scholar
  12. Durrant HJ, Ratcliffe NA, Hipkin CR, Aspan A, Soderhall K. Purification of the prophenoloxidase enzyme from hemocytes of the cockroach Blaberus discoidalis. Biochem. J. 1993; 289: 87–91.PubMedGoogle Scholar
  13. Fujimoto K, Masuda K, Asada N, Ohnishi E. Purification and characterization of prophenoloxidase from the pupae of Drosophila melanogaster. J. Biochem (Tokyo). 1993; 113: 285–291.Google Scholar
  14. Fujimoto K, Okino N, Kawabata SI, Iwanaga S, Ohnishi E. Nucleotide sequence of the eDNA encoding the proenzyme of phenoloxidase A of Drosophila melanogaster. Proc. Natl. Acad. Sci USA. 1995; 92: 7769–7773.PubMedCrossRefGoogle Scholar
  15. Gillespie JP, Kanost MR, Trenczek T. Biological mediators of insect immunity. Ann. Rev. Entomol. 1997; 42: 611–643.CrossRefGoogle Scholar
  16. Hall M, Scott M, Sugumaran M, Soderhall K, Law JH. Proenzyme of Manduca sexta phenoloxidase: Purification, activation, substrate specificity of the active enzyme and molecular cloning. Proc. Natl. Acad. Sci USA. 1995; 92: 7764–7768.PubMedCrossRefGoogle Scholar
  17. Hara T, Miyoshi T, Funatsu M. Comparative studies on larval and pupal phenoloxidases of the housefly, Musca domestica. Comp. Biochem. Physiol. 1993; 106B: 287–292.Google Scholar
  18. Heyneman RA. Final purification of a latent phenoloxidase with mono-and diphenoloxidase from Tenebrio molitor. Biochem. Biophys. Res. Commun. 1965; 21: 162–169.PubMedCrossRefGoogle Scholar
  19. Jiang H, Wang Y, Ma C, Kanost MR. Subunit composition of prophenoloxidase from Manduca sexta: Molecular cloning of subunit ProPo-P1. Insect Biochem. Mol. Biol. 1997a; 27, 835–850.CrossRefGoogle Scholar
  20. Jiang H, Wang Y, Korochkina SE, Benes H, Kanost MR. Molecular cloning of cDNAs for two prophenoloxidases subunits from the Malaria vector, Anopheles Gambiae. Insect Biochem. Mol. Biol. 1997b; 27: 693–699.CrossRefGoogle Scholar
  21. Kawabata T, Yasuhara Y, Ochiai M, Matsuura S, Ashida M. Molecular cloning of insect prophenoloxidase: A copper containing protein homologous to arthropod hemocyanin. Proc. Natl. Acad. Sci. USA. 1995; 92: 7774–7778.PubMedCrossRefGoogle Scholar
  22. Kwon TH, Lee SY, Lee JH, Choi JS, Kawabata SI, Iwanaga S, Lee BL. Purification and characterization of prophenoloxidase from the hemolymph of coleopteran insect, Holotrichia diomphlia. Molecules & Cells 1997; 7: 90–97.Google Scholar
  23. Lai-Fook J. The repair of wounds in the integument of insects. J. Insect Physiol. 1966; 12:195–226. CrossRefGoogle Scholar
  24. Lee WJ, Ahmed A, Torre AD, Kobayashi A, Ashida M, Brey PT. Molecular cloning and chromosomal localization of a prophenoloxidase eDNA from the malaria vector Anapheles gambiae. Insect Mol. Biol. 1998; 7:41–50.PubMedCrossRefGoogle Scholar
  25. Lewis HW, Lewis HS. Genetic regulation of dopa oxidase activity in Drosophila. Ann. N.Y. Acad. Sci. 1963; 100: 827–839.Google Scholar
  26. Maddison WP, Maddison DR. MACLADE, Version 3.04. Sunderland Mass, USA. SinauerAssociates, Inc. Publishers. 1993.Google Scholar
  27. Muller HM, Dimopoulos G, Blass C, Kafatos FC. A hemocyte-like cell line established from malaria vector Anopheles gambiae expresses six prophenoloxidase genes. J. Biol. Chem. 1999; 274: 11727–11735.PubMedCrossRefGoogle Scholar
  28. Nappi AJ, Sugumaran M. “Some biochemical aspects of eumelanin formation in insect immunity”. In: Insect Immunity. Pathak J. P. N. cd. New Delhi, India. Oxford & IBH Publishing Co. 1993. pp. 131–148.Google Scholar
  29. Pau RN, Eagles PAM. The isolation of o-diphenoloxidase from the third instar larvae of blowfly, Calliphora erythocephala. Biochem. J. 1975; 149: 707–712.PubMedGoogle Scholar
  30. Park DS, Shin W, Kim MG, Park SS, Lee WJ, Brey PT, Park HY. Isolation and characterization of the eDNA encoding the prophenoloxidase of Fall webworm, Hyphantria cunea. Insect Biochem. Mol. Biol. 1997; 27: 983–992.PubMedCrossRefGoogle Scholar
  31. Pentz ES, Wright TR. Dmsophila melanogaster diphenoloxidase A2: Gene structure and homology with the mouse mast-cell turn-transplantation antigen, P91A. Gene 1991; 22: 239–242.CrossRefGoogle Scholar
  32. Perotti ME, Bairati A. Ultrastructure of the melanotic masses in two tumorous strains of Drosophila melanogaster (tuB3 and Freckled). J. Invert. Pathol. 1968; 10: 122–138.CrossRefGoogle Scholar
  33. Ricketts D, Sugumaran M. 1,2-dehydro-N-b-alanyldopamine as a new intermediate in insect cuticular sclerotization. J. Biol. Chem. 1994; 269: 22217–22221.PubMedGoogle Scholar
  34. Saul SJ, Sugumaran M. A novel quinone: quinone methide isomerase generates quinone methides in insect cuticle. F. E. B. S. Lett. 1988; 237: 155–158.CrossRefGoogle Scholar
  35. Saul SJ, Sugumaran M. Characterization of a new enzyme system that desaturates the side chain of Nacetyldopamine. F.E.B.S. Lett. 1989a; 251: 69–73.CrossRefGoogle Scholar
  36. Saul SJ, Sugumaran M. N-Acetyldopamine quinone methide/1,2-dehydro-N-acetyldopamine tautomerase - A new enzyme involved in sclerotization of insect cuticle. F.E.B.S. Lett. 1989b; 255: 340–344.CrossRefGoogle Scholar
  37. Saul SJ, Sugumaran M. 4-alkyl-o-quinone/2-hydroxy-p-quinone methide isomerase from the larvae hemolymph of Sarcophaga bullata. I. Purification and characterization of enzyme catalyzed reaction. J. Biol. Chem. 1990; 265: 16992–16999.PubMedGoogle Scholar
  38. Seybold WD, Meltzer PS, Mitchell HK. Phenoloxidase activation in Drosophila: A cascade of reactions. Biochem. Genetics. 1975; 13: 85–108.Google Scholar
  39. Söderhäll K, Aspán A, Duvic B. The ProPO system and associated proteins. Role in cellular communication in arthropods. Res. Immunol. 1990; 141: 896–907.PubMedGoogle Scholar
  40. Sugumaran M. Molecular mechanisms of mammalian melanogenesis - comparison with insect cuticular sclerotization. F. E. B. S. Lett. 1991; 293: 4–10.CrossRefGoogle Scholar
  41. Sugumaran M. “Role of insect cuticle in immunity”. In New Directions in Invertebrate Immunology. Söderhäll, K., Iwanaga, S. and Vastha, G. eds. Fair Haven, NJ. SOS Publications. 1996. pp. 355–374.Google Scholar
  42. Sugumaran M. Unified mechanism for sclerotization of insect cuticle. Adv. Insect Physiol. 1998; 27: 229–334.CrossRefGoogle Scholar
  43. Sugumaran M, Kanost M. “Regulation of insect hemolymph phenoloxidases”. In Parasites and pathogens Beckage, N. E., Thompson, S. N., & Frederick, B. A. eds. San Diego, Academic Press. 1993; Vol. I. Parasites. pp. 317–342.CrossRefGoogle Scholar
  44. Stoltzfus A, Logsdon JM, Palmer JD, Doolittle WF. Intron “sliding” and the diversity of intron positions. Proc. Natl. Acad. Sci. USA 1997; 94: 10739–10744.PubMedCrossRefGoogle Scholar
  45. Thompson JD, Higgins DG, Gibson TJ. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions-specific gap penalties and weight matrix choice. Nucleic Acid Res. 1994; 22: 4673–4680.PubMedCrossRefGoogle Scholar
  46. Yasuhara Y, Koizumi Y, Katagiri C, Ashida M. Reexamination of properties of prophenoloxidase isolated from larval hemolymph of the silkworm, Bombyx mori. Arch. Insect Biochem. Physiol. 1995; 32: 14–23.Google Scholar

Copyright information

© Springer Science+Business Media New York 2001

Authors and Affiliations

  • Michael R. Chase
    • 1
  • Manickam Sugumaran
    • 1
  1. 1.Department of BiologyUniversity of Massachusetts - BostonBoston

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