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Evidence for proteins involved in prophenoloxidase cascade Eisenia fetida earthworms

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Abstract

The prophenoloxidase cascade represents one of the most important defense mechanisms in many invertebrates. Following the recognition of microbial saccharides by pattern recognition molecules, proteinases cleave inactive prophenoloxidase to its active form, phenoloxidase. Phenoloxidase is a key enzyme responsible for the catalysis of the melanization reaction. Final product melanin is involved in wound healing and immune responses. Prophenoloxidase cascade has been widely described in arthropods; data in other invertebrate groups are less frequent. Here we show detectable phenoloxidase activity in 90-kDa fraction of the coelomic fluid of earthworms Eisenia fetida. Amino acid sequencing of peptides from the active fraction revealed a partial homology with invertebrate phenoloxidases and hemocyanins. Moreover, the level of phenoloxidase activity is lower and the activation slower as compared to other invertebrates.

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Abbreviations

CF:

Coelomic fluid

LBSS:

Lumbricus balanced salt solution

PBS:

Phosphate buffered solution

proPO:

Prophenoloxidase

PO:

Phenoloxidase

l-DOPA:

l-β-3,4-Dihydroxyphenylalanine

ppA:

Prophenoloxidase-activating enzyme

CPC:

Cetylpyridium chloride

Hc:

Hemocyanin

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Acknowledgements

This work was supported by the Czech Science Foundation (310/04/0806; B500200613), Institutional Research Concept (AVOZ50200510) and a bilateral international scientific and technological cooperation grant of the Ministry of the Flemish Community (BOF-BWS 03/06) and was performed within the frames of an Interuniversity Attraction Pole Program. Authors are grateful to Prof. Söderhäll, Uppsala, for providing probe containing the gene coding for PO in crayfish P. leniusculus. All experiments comply with the current laws of the Czech Republic and Belgium.

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Authors and Affiliations

  1. Department of Immunology, Institute of Microbiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20, Prague 4, Czech Republic

    Petra Procházková, Marcela Šilerová, Radka Josková & Martin Bilej

  2. Department of Cellular and Molecular Interactions, Vlaams Interuniversitair Instituut voor Biotechnologie, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium

    Benoit Stijlemans, Alain Beschin & Patrick De Baetselier

  3. Department of Biogenesis and Biotechnology of Natural Compounds, Institute of Microbiology, Academy of Sciences of the Czech Republic, Videnska 1083, 142 20, Prague 4, Czech Republic

    Petr Halada

  4. Facultés Universitaires Notre-Dame de la Paix, Unité de Recherche en Biologie Cellulaire, Spectrométrie de Masse, rue de Bruxelles 61, 5000, Namur, Belgium

    Marc Dieu

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  1. Petra Procházková
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  2. Marcela Šilerová
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  3. Benoit Stijlemans
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  9. Martin Bilej
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Correspondence to Petra Procházková.

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Communicated by G. Heldmaier

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Procházková, P., Šilerová, M., Stijlemans, B. et al. Evidence for proteins involved in prophenoloxidase cascade Eisenia fetida earthworms. J Comp Physiol B 176, 581–587 (2006). https://doi.org/10.1007/s00360-006-0081-z

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  • DOI: https://doi.org/10.1007/s00360-006-0081-z

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