Skip to main content
SpringerLink
Log in

Prophenol Oxidase A3 in Drosophila melanogaster: Activation and the PCR-Based cDNA Sequence

  • Published:
Biochemical Genetics Aims and scope Submit manuscript

Abstract

Phenol oxidase exists in Drosophila hemolymph as a prophenol oxidase, A1 and A3, that is activated in vivo with a native activating system, AMM-1, by limited proteolysis with time. The polypeptide in purified prophenol oxidase A3 has a molecular weight of approximately 77,000 Da. A PCR-based cDNA sequence coding A3 has 2501 bp encoding an open reading frame of 682 amino acid residues. The potential copper-binding sites, from Trp-196 to Tyr-245, and from Asn-366 to Phe-421, are highly homologous to the corresponding sites in other invertebrates. The availability of prophenol oxidase cDNA should be useful in revealing the biochemical differences between A1 and A3 isoforms in Drosophila melanogaster that are refractory or unable to activate prophenol oxidase.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Asada, N. (1997). Genetic variants affecting phenol oxidase activity in Drosophila melanogaster. Biochem. Genet. 35:41.

    PubMed  Google Scholar 

  • Asada, N. (1998). Reversible activation of prophenoloxidase with 2–propanol in Drosophila melanogaster. J. Exp. Zool. 282:28.

    PubMed  Google Scholar 

  • Asada, N., Fujimoto, K., Tanaka, M., & Ohnishi, E. (1993) Genetic polymorphism of prophenoloxidase A1 in Drosophila melanogaster. Jpn. J. Genet. 68:219.

    PubMed  Google Scholar 

  • Asada, N., & Sezaki, H. (1999) Properties of phenoloxidase generated from prophenoloxidase with 2–propanol and the natural activator in Drosophila melanogaster. Biochem. Genet. 37: 149.

    PubMed  Google Scholar 

  • Ashida, M., & Brey, P. T. (1997). Recent advances in prophenoloxidase research. In Brey, P. T., & Hultmark, D. (eds.), Molecular Mechanisms of the Insect Immune Response, Chapman and Hall, London, p. 135.

    Google Scholar 

  • Ashida, M., & Yamazaki, H. I. (1990). Biochemistry of the phenoloxidase system in insects: With special reference to its activation. In Ohnishi, E., & Ishizaki, H. (eds.), Molting and Metamorphosis, Japan Scientific Societies Press, Tokyo, p. 239.

    Google Scholar 

  • Aspan, A., Huang, T. S., Cerenius, L., & Söderhäll, K. (1995). cDNA cloning of a prophenoloxidase from the freshwater crayfish Pacifastacus leniusculus and its activation. Proc. Natl. Acad. Sci. U.S.A. 92:936.

    Google Scholar 

  • Chase, M. R., Raina, K., Bruno, J., & Sugumaran, M. (2000). Purification, characterization and molecular cloning of prophenol oxidase from Sarcophaga bullata. Insect Biochem. Mol. Biol. 30:853.

    Google Scholar 

  • Cherqui, A., Duvic, B., & Breherin, M. (1996). Purification and characterization of prophenoloxidase from the hemolymph of Locusta migratoria. Arch. Insect Biochem. Physiol. 32:225.

    Google Scholar 

  • Chomchzynsky, P., & Sacchi, N. (1987). Modified isolation for Manduca epidermis. Anal. Biochem. 162:156.

    PubMed  Google Scholar 

  • Chosa, N., Fukumitsu, T., Fujimoto, K., & Ohnishi, E. (1997). Activation of prophenol oxidase A1 by an activating enzyme in Drosophila melanogaster. Insect Biochem. Mol. Biol. 27:61.

    PubMed  Google Scholar 

  • Cui, L., Luckhart, S., & Rosenberg, R. (2000). Molecular characterization of prophenoloxidase cDNA from the malaria mosquito Anopheles stephensi. Insect Mol. Biol. 9:127.

    PubMed  Google Scholar 

  • Fujimoto, K., Masuda, K., Asada, N., & Ohnishi, E. (1993). Purification and characterization of prophenoloxidases from pupae of Drosophila melanogaster. J. Biochem. 113:285.

    PubMed  Google Scholar 

  • Fujimoto, K., Okino, N., Kawabata, S., Iwanaga, S., & Ohnishi, E. (1995). Nucleotide sequence of the cDNA encoding the proenzyme of phenol oxidase A1 of Drosophila melanogaster. Proc. Natl. Acad. Sci. U.S.A. 92:7769.

    PubMed  Google Scholar 

  • Hall, M., Scott, T., Sugumaran, M., Söderhäll, K., & Law, J. H. (1995). Proenzyme of Manduca sexta phenol oxidase: Purification, activation, substrate specificity of the active enzyme, and molecular cloning. Proc. Natl. Acad. Sci. U.S.A. 92:7764.

    PubMed  Google Scholar 

  • Hirano, H., Komatsu, S., Takakura, H., Sakiyama, F., & Tsunasawa, S. (1992). Deblocking and subsequence analysis of Na-blocked proteins electroblotted onto PVDF membrane. J. Biochem. 111:754.

    PubMed  Google Scholar 

  • Jiang, H., Wang, Y., Ma, C., & Kanost, M. R. (1997). Subunit composition of pro-phenol oxidase from Manduca sexta: Molecular cloning of subunit proPO-P1. Insect Biochem. Mol. Biol. 27: 835.

    PubMed  Google Scholar 

  • Kawabata, T., Yasuhara, Y., Ochiai, M., Matsuura, S., & Ashida, M. (1995). Molecular cloning of insect pro-phenol oxidase: A copper-containing protein homologous to arthropod hemocyanin. Proc. Natl. Acad. Sci. U.S.A. 92:7774.

    PubMed  Google Scholar 

  • Laemmli, U. K. (1970). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680.

    PubMed  Google Scholar 

  • Lee, H. S., Cho, M. Y., Lee, K. M., Kwon, T. M., Homma, K., Nutria, S., & Lee, B. L. (1999). The pro-phenol oxidase of coleopteran insect, Tenebrio molitor, larvae was activated during cell climp/cell adhesion if insect cellular defense reactions. FEBS Lett. 444:255.

    PubMed  Google Scholar 

  • Ohnishi, E. (1953). Tyrosinase activity during puparium formation in Drosophila melanogaster. Jpn. J. Zool. 11:69.

    Google Scholar 

  • Park, D. S., Shin, S. W., Kim, M. G., Park, S. S., Lee, W. J., Brey, P. T., & Park, H. Y. (1997). Isolation and characterization of the cDNA encoding the prophenoloxidase of fall webworm, Hypantria cunea. Insect Biochem. Mol. Biol. 27:983.

    PubMed  Google Scholar 

  • Rizki, T. M., Rizki, R. M., & Bellotti, R. A. (1985). Genetics of Drosophila phenoloxidase. Mol. Gen. Genet. 201:7.

    Google Scholar 

  • Sambrook, J., Fritsh, E. F., & Maniatis, T. (1989). Molecular Cloning: A Laboratory Manual, 2nd edn., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY.

    Google Scholar 

  • Sanger, F., Nicklen, S., & Coulson, A. R. (1977). DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. U.S.A. 74:5463.

    PubMed  Google Scholar 

  • Sezaki, H., Kawamoto, N., & Asada, N. (2001). Effect of ionic concentration on the higher-order structure of prophenol oxidase in Drosophila melanogaster. Biochem. Genet. 39:50.

    Google Scholar 

  • Söderhäll, K., Cerenius, L., & Johanson, M. W. (1996). The prophenoloxidase activating system in invertebrates. In Soderhall, K., Iwanaga, S., & Vasta, G. R. (eds.), New Directions in Invertebrate Immunology, SOS Publications, Fair Haven, NJ.

    Google Scholar 

  • Sugumaran, M. (1996). Role of insect cuticle in immunity. In Soderhall, K., Iwanaga, S., & Vasta, G. R. (eds.), New Directions in Invertebrate Immunology, SOS Publications, Fair Haven, NJ.

    Google Scholar 

  • Tsunasawa, S., Takakura, H., & Sakiyama, F. (1990). The amino acid sequence of S-antigen: N-terminus and uvenitogenic peptides. J. Protein Chem. 9:256.

    Google Scholar 

  • Yonemura, M., Kasatani, K., Asada, N., & Ohnishi, E. (1991). Involvement of a serine protease in the activation of prophenoloxidase in Drosophila melanogaster. Zool. Sci. 8:865.

    Google Scholar 

Download references

Authors
  1. Nobuhiko Asada
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Genta Yokoyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nobuko Kawamoto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shigemi Norioka
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takashi Hatta
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Asada, N., Yokoyama, G., Kawamoto, N. et al. Prophenol Oxidase A3 in Drosophila melanogaster: Activation and the PCR-Based cDNA Sequence. Biochem Genet 41, 151–163 (2003). https://doi.org/10.1023/A:1023325610300

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1023325610300

Search

Navigation