Control Mechanisms of the Prophenoloxidase Cascade
Part of the
Advances in Experimental Medicine and Biology
book series (AEMB, volume 484)
Insects do not possess innnunoglobulins and other complicated immune proteins found in higher animals. However, they are equipped with a highly effective defense arsenal to protect themselves from invading parasites and other intruders. The hard armor like exoskeleton of insects affords the first line of defense by providing a physical barrier against the foreign organisms (Ashida & Brey, 1995; Sugumaran 1996,1998a). Once the cuticular barrier is surcumvented, the organisms entering the soft body of insects face a plethora of cellular and humoral host defense reactions. The response to the presence of non-self matter includes, phagocytocysis, nodule formation, encapsulation, melanization, and synthesis and secretion of antibacterial proteins (Hoffman et al., 1999; Gillespie et al., 1997; Södärhall et al., 1990; Sugumaran 1996). Organisms too large to be phagocytosed are usually found encapsulated and often melanized in the hemolymph of insects. Therefore, phenoloxidase (PO), the enzyme responsible for the biosynthesis of melanin is considered as an important component of insects’ immune system (Ashida & Yamazaki, 1990; Gillespie et a1.,1997; Sugumaran & Kanost 1993; Nappi & Sugumaran 1993; Soderhall et al., 1990). Interestingly, PO is not only involved in defense reactions, but also in a few other physiologically important and relevant processes. Cuticular sclerotization which is essential to the survival of all insects is initiated by PO (Sugumaran 1998a). The quinonoid products formed, are acted upon by other cuticular enzymes such as quinone isomerase and quinone methide isomerase. These enzymes in association with PO, generate a number of reactive intermediates that crosslinking structural proteins and chitin. These reactions render the cuticular proteins insoluble and eventually cause the hardening of cuticle. In the last process, viz., wound healing, massive amounts of hemolymph loss is partly prevented by the action of PO which rapidly deposits melanin pigment at the wounding site (Ashida & Brey 1995; Lai-Fook, 1966; Sugumaran 1996). In addition, PO generated quinones being cytotoxic, may play a critical role in killing opportunistically invading microorganisms at the wound site (Nappi & Sugumaran 1993; Sugumaran 1996).
KeywordsPhenol Oxidase Cuticular Protein Phenoloxidase Activity Insect Immunity Prophenoloxidase Activation
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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:
Ashida M. Purification and characterization ofprephenoloxidase from hemolymph of the silkworm, Bombyx mori.
Arch. Biochem. Biophys. 1971; 144:
Ashida M, Brey P. Role of the integument in insect defense: Prophenoloxidase cascade in the cuticular matrix. Proc. Natl. Acad. Sci. USA. 1995; 92:
Ashida M, Dohke K. Activation of pro-phenoloxidase by the activating enzyme of the silkworm, Bombyx mori
. Insect. Biochem. 1980; 10: 37–47.Google Scholar
Ashida M, Sasaki T. A target protease of serpins in insect hemolymph. Insect. Biochem. Mol. Biol. 1994; 24:
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
Ashida M, Yoshida H. Limited proteolysis of prophenoloxidase during activation by microbial products in insect plasma and effect of phenoloxidase on electrophoretic mobilities of plasma proteins. Insect Biochem. 1988; 18:
Aso Y, Kramer KJ, Hopkins TL, Lookhart GL. Characterization of hemolymph protyrosinase and a cuticular activator from Manduca sexta
(L). Insect Biochem. 1985; 15:
Aso Y, Yamashita T, Meno K, Murakami M. Inhibition of prophenoloxidase-activating enzyme from Bombyx mori
by endogenous chymotyrpsin inhibitors. Biochem. Mol. Biol. Int. 1994; 33:
Beck G, Cardinale S, Wang L, Reiner M, Sugumaran M. Characterization of a defense complex consisting of interleukin 1 and phenoloxidase from the hemolymph of the tobacco hornworm, Manduca sexta.
J. Biol. Chem. 1996;. 271:
Boigegrain RA, Mattras H, Brehelin M, Paroutaud P, Coletti-Previero MA. Insect immunity: Two proteinase inhibitors from hemolymph of Locusta migratoria
Biochem. Biophys. Res. Commun. 1992: 189
, 790–793.Google Scholar
Brehelin M, Boigegrain RA, Drif L, Coletti-Previero MA. Purification of a protease inhibitor which controls prophenoloxidase activation in hemolymph of Locusta migratoria
(insecta). Biochem. Biophys. Res. Commun. 1991: 179
, 841–846.Google Scholar
Chase MR, Sugumaran M. Genomic and cDNA sequences of prophenoloxidases from Drosophila melanogaster.
This volume, page 349–362. (2000)Google Scholar
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
Cherqui A, Duvic B, Brehelin M. Purification and characterization of prophenoloxidase from the hemolymph of Locusta migratoria.
Arch. Insect Biochem. Physiol. 1996; 32:
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:
Chosa N, Fukumitsu T
, Fujimoto K, Ohnishi E. Activation of pro-phenoloxidase A, by an activating enzyme in Drosophila melanogaster
. Insect Biochem. Mol. Biol. 1997; 27:
Daquinag AC, Nakamura S, Takao T, Shimonishi Y, Tsukamoto T. Primary structure of a potent inhibitor of phenol oxidase from Musca Domestica.
Proc. Natl. Acad. Sei. USA. 1995: 92
, 2964–2968.CrossRefGoogle Scholar
Daquinag AC, Sato T. Kodo H, Takao T, Fukuda M, Shimonishi Y, Tsukamoto T. A novel endogenous inhibitor of phenol oxidase from Musca Domestica
has a cystine motif commonly found in snail and spider toxins. Biochemistry. 1999: 38, 2179–2188.PubMedCrossRefGoogle Scholar
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
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
Fujimoto K, Okino N, Kawabata SI, Iwanaga S, Ohnishi E. Nucleotide sequence of the cDNA encoding the proenzyme of phenoloxidase Ai
of Drosophila melanogaster.
Proc. Natl. Acad. Sci USA. 1995; 92:
Gillespie JP, Kanost MR, Trenczek T. Biological mediators of insect immunity. Ann. Rev. Entornol. 1997; 42:
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:
Hara T, Miyoshi T, Funatsu M. Comparative studies on larval and pupal phenoloxidases of the housefly, Musca domestica.
Comp. Biochem. Physiol. 1993; 106B:
Heyneman RA. Final purification of a latent phenoloxidase with mono-and diphenoloxidase from Tenebrio molitor.
Biochem. Biophys. Res. Commun. 1965; 21: 162–169.CrossRefGoogle Scholar
Hoffmann JA, Kafatos FC, Janeway CA, Ezekowitz RAB. Phylogenetic perspectives in innate immunity. Science 1999; 284: 1313–1318.PubMedCrossRefGoogle Scholar
Jiang H, Kanost MR. Characterization and functional analysis of twelve naturally occurring reactive site variants of serpins-1 from Manduca sexta.
J. Biol. Chem. 1997; 272:
Jiang H, Wang Y, Huang Y, Mulnix AB, Kadel J, Cole K, Kanost MR. Organization of serpin gene 1 from Manduca sexta.
Evolution of a family of alternate exons encoding the reactive site loop. J. Biol. Chem. 1996; 271:
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
Jiang H, Wang Y, Korochkina SE, Benes H, Kanost MR. Molecular cloning of cDNAs for two prophenoloxidases subunits from the Malaria vector, Anopheles Gambia
e. Insect Biochem. Mol. Biol. 1997b; 27:
Jiang H, Wang Y, Kanost MR. Prophenoloxidase activating proteinase from an insect, Manduca sexta.
A bacterial inducible protein similar to Drosophila
ester. Proc. Natl. Acad. Sci USA. 1998; 95:
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:
Kopácek P, Weise C, Gotz P. The prophenoloxidase from the wax moth Galleria mellonella:
Purification and characterization of proenzyme. Insect Biochem. Mol. Biol. 1995; 25:
Kramer KJ, Hopkins TL. Tyrosine metabolism for insect cuticle tanning. Arch. Insect Biochem. Physiol. 1987; 6:
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:
Lai-Fook J. The repair of wounds in the integument of insects. J. Insect Physiol. 1966; 12:195–226.
Lee WJ, Ahmed A, Torre AD, Kobayashi A, Ashida M, Brey PT. Molecular cloning and chromosomal localization of a prophenoloxidase cDNA from the malaria vector Anopheles gambiae
. Insect Mol. Biol. 1998a; 7:41–50.CrossRefGoogle Scholar
Lee SY, Kwon TH, Hyun JH, Choi JS, Kawabata SI, Iwanaga S, Lee BL. In vitro
activation of prophenoloxidase by two kinds of prophenoloxidase activating factors isolated from hemolymph of coleopteran, Holotrichia diomphlia
larvae. Eur. J. Biochem. 1998b; 254:
Lee SY, Cho MY, Hyun JH, Lee KM, Homma K, Natori S, Kawabata SI, Iwanaga S, Lee BL. Molecular cloning of cDNA for prophenoloxidase activating factor I, a serine protease is induced by lipopolysaccharide orb-1,3-glucan in coleopteran insect Holotrichia diomphlia.
larvae. Eur. J. Biochem. 1998e; 257:
Muller HM, Dimopoulos G, Blass C, Kafatos FC. Ahemocyte-like cell line established from malaria vector Anopheles gambiae
expresses six prophenoloxidase genes. J. Biol. Chem. 1999; 274:
Nappi AJ, Sugumaran M. “Some biochemical aspects of eumelanin formation in insect immunity”. In: Insect Immunity. Pathak J. P. N. ed. New Delhi, India. Oxford & IBH Publishing Co. 1993. pp. 131–148.Google Scholar
Park DS, Shin W, Kim MG, Park SS, Lee WJ, Brey PT, Park HY. Isolation and characterization of the cDNA encoding the prophenoloxidase of Fall webworm, Hyphantria cunea
. Insect Biochem. Mol. Biol. 1997; 27:
Pau RN, Eagles PAM. The isolation ofo-diphcnoloxidase from the third instar larvae of blowfly, Calliphora erythocephala.
Biochem. J. 1975; 149:
Ramesh N, Sugumaran M, Mole JE. Purification and characterization of two trypsin inhibitors from the hemolymph of Manduca sexta
larvae. J. Biol. Chem. 1988; 263:
, Horii A, Ochiai M, Ashida M. Prophenoloxidase activating enzyme of silkworm Bombyx mori
-Purification, characterization and cDNA cloning. J. Biol. Chem. 1999: 274:
Saul SJ, Sugumaran M. Protease inhibitor controls prophenoloxidase activation in Manduca sexta.
F. E. B. S. Lett. 1986; 208:
Saul SJ, Sugumaran M.
Protease mediated prophenoloxidase activation in the hemolymph of the tobacco homworm, Manduca sexta
. Arch. Insect Biochem. Physiol. 1987; 5:
Saul SJ, Sugumaran M. Prophenoloxidase activation in the hemolymph ofSarcophaga bullata
larvae. Arch. Insect Biochem. Physiol. 1988; 7: 91–103.Google Scholar
Saul SJ, 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; 11:
Söderhäll K, Aspán A, Duvic B. The ProPO system and associated proteins. Role in cellular communication in arthropods. Res. Immunol. 1990; 141:
Srere PA. Complexes of sequential metabolic enzymes. Ann. Rev. Biochem. (1987). 56:
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
Sugumaran M. Unified mechanism for sclerotization of insect cuticle. Adv. Insect Physiol. 1998a; 27:
“characterization ofphenoloxidasc complexes”. In Techniques in Insect Immunology.
Wiesner A, Dunphy GB, Marmaras VJ, Morishima I, Sugumaran M, Yamakawa M. eds. Fair Haven, NJ. SOS Publications. 1998. pp. 205–215.Google Scholar
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
Sugumaran M, Nellaiappan K. On the latency and nature of phenoloxidase present in the left colleterial gland of the cockroach, Periplaneta americana
. Arch. Insect Biochem. Physiol. 1990; 15:
Sugumaran M, Nellaiappan K. Characterization of a new phenoloxidase inhibitor from the cuticle of Manduca sexta
. Biochem. Biophys. Res. Commun. 2000; 268:
Sugumaran M, Nellaiappan K. Lysolecithin - A potent activator of prophenoloxidase from the hemolymph of the lobster, Homarus americanas
. Biochem. Biophys. Res. Commun. 1991; 176:
Sugumaran M, Saul SJ, Ramesh N. Endogenous protease inhibitors prevent undesired activation of prophenolase in insect hemolymph. Biochem. Biophys. Res. Commun. 1985; 132:
Sugumaran M, Nellaiappan K, Scott T, Amarathunga C. Complex formation between mushroom tyrosinasc and Manduca
dopachrome isomerase. Pigment Cell. Res. 1995; 8:
Sugumaran M, Nellaiappan K, Amarathunga C, Cardinale S, and Scott T. Insect Melanogenesis. III Metabolon formation in the melanogenic pathway—Regulation of phenoloxidase activity by endogenous dopachrome isomerase (Decarboxylating) from Manduca sexta
. Arch. Biochem. Biophys.2000; 378:
Tsukamoto, T., Ichimaru, Y., Kanegae, N., Watanabe, K., Yamaura I., Katasura, Y. and Funatsu, M. Identification and isolation of endogenous insect phenoloxidase inhibitors. Biochem. Biophys. Res. Commun.
, 86–92.PubMedCrossRefGoogle Scholar
Yasuhara Y, Koizumi Y, Katagiri C, Ashida M. Reexamination of properties of prophenoloxidase isolated from larval hemolymph of the silkworm, Bombyx mori.
Arch. Biochem. Biophys. 1995; 320:
, Ashida M. Microbial activation of two scrinc enzymes and prophenoloxidase in the plasma fraction of hemolymph of the silkworm, Bombyx mori.
Insect Biochem. Mol. Biol. 1986; 16:
© Springer Science+Business Media New York 2001