Skip to main content

Advertisement

Springer Nature Link
Log in

Identification and characterization of proteases from skin mucus of tambacu, a Neotropical hybrid fish

  • Published:
Fish Physiology and Biochemistry Aims and scope Submit manuscript

Abstract

Skin secretions of fishes constitute a rich source of proteins with a broad spectrum of antimicrobial properties. We report here the characterization of proteases from skin mucus of tambacu, an economically important Neotropical hybrid fish. The effects of pH on the proteolytic activities of the mucus acting on various substracts – hemoglobin, casein, bovine serum albumin (BSA) and ovalbumin (OVA) – were tested. Optimal pH values for protease activity on hemoglobin were 4.5 and 8.5, on casein, 8.5, on BSA, 5.0 and 7.5, and on ovalbumin, 4.5 and 6.5. The proteolytic activity was inhibited on all of these substrates in the presence of specific inhibitors: caseinolytic activity was inhibited by inhibitors of serine and metalloproteases; hemoglobinolytic activity was inhibited by serine, aspartic and metalloproteases inhibitors; albuminolytic activity on BSA was inhibited by serine and aspartic proteases inhibitors, and on ovalbumin, by cysteine and aspartic proteases inhibitors. Gelatin zymography revealed that the skin mucus of tambacu consisted primarily of serine and metalloproteases. Hemoglobin zymography showed one proteolytic band inhibited by EDTA, whereas casein zymography showed two proteases inhibited by serine proteases inhibitors. We were able to identify all classes of proteases in the mucus from the skin of tambacu. These, and these results suggest that the proteolytic activities of the skin mucus of fish may play an important role in the defense against microorganisms and ectoparasites.

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

Access this article

Log in via an institution

Subscribe and save

Springer+
from $39.99 /Month
  • Starting from 10 chapters or articles per month
  • Access and download chapters and articles from more than 300k books and 2,500 journals
  • Cancel anytime
View plans

Buy Now

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Explore related subjects

Discover the latest articles and news from researchers in related subjects, suggested using machine learning.

Abbreviations

ACA:

6-Amino-caproic acid

CEPTA/IBAMA:

Centro de Pesquisas de Peixes Neotropicais

E-64:

l-trans-3-Carboxyoxiran-2-carbonyl-l-leucylagmatine

EDTA:

Ethylenediaminetetraacetic acid

pHMB:

p-Hydroxy-mercurybenzoate

PMSF:

Phenylmethylsulfonyl fluoride

TLCK:

N-Tosyl-lysine chloromethyl ketone

TPCK:

N-Tosyl-l-phenylalanine chloromethyl ketone

References

  • Aranishi F (1999) Lysis of pathogenic bacteria by epidermal cathepsins L and B in the Japanese eel. Fish Physiol Biochem 20:37–41

    Article  CAS  Google Scholar 

  • Aranishi F, Nakane M (1997a) Epidermal proteinases in the European eel. Physiol Zool 70:563–570

    Article  CAS  Google Scholar 

  • Aranishi F, Nakane M (1997b) Epidermal proteinases of the Japanese eel. Fish Physiol Biochem 16:471–478

    Article  CAS  Google Scholar 

  • Aranishi F, Nakane M (1998a) Epidermal proteases of American eel. J Aquat Anim Health 10:35–42

    Article  Google Scholar 

  • Aranishi F, Nakane M (1998b) Epidermal proteases of Japanese eel. Fish Physiol Biochem 16:471–478

    Article  Google Scholar 

  • Bernardino G, Mendonça JOJ, Ribeiro LP, Alcântara RCGV, Ferrari A, Fijan N (1986) First reproduction of tambacu – a Colossoma hybrid. In: Síntese dos trabalhos realizados com espécies do gênero Colossoma. CEPTA, Pirassununga, pp 11–12

  • Buchmann K, Bresciani J (1999) Rainbow trout leucocytes activity: influence on the ectoparasitic monogenean Gyrodactylus deriavini. Dis Aquat Organ 35:13–22

    PubMed  CAS  Google Scholar 

  • Calcagnotto D, Almeida-Toledo LF, Bernardino G, Toledo Filho SA (1999) Biochemical genetic characterization of F1 reciprocal hybrids between pacu (Piaractus mesopotamicus) and tambaqui (Colossoma macropomum) reared in Brazil. Aquaculture 174:51–57

    Article  CAS  Google Scholar 

  • Cho JH, Park IY, Kim HS, Lee WT, Kim MS, Kim SC (2002) Cathepsin D produces antimicrobial peptide parasin I from histone H2A in the skin mucosa of fish. FASEB J 16:429–431

    PubMed  CAS  Google Scholar 

  • Fast MD, Sims DE, Burka JF, Mustafa A, Ross NW (2002) Skin morphology and humoral non-specific defence parameters of mucus and plasma in rainbow trout, coho and Atlantic salmon. Comp Biochem Physiol 132:645–657

    CAS  Google Scholar 

  • Goulding M (1981) In: Dumont HJ (ed) Developments in hydrobiology. Dr. W. Junk Publ, The Hague, p 137

  • Hernández R (ed) (1989) Cultivo de colossoma. Editora Guadalupe, Bogota

  • Hjelmeland K (1983) Skin mucus protease from rainbow trout, Salmo gairdneri Richardson, and its biological significance. J Fish Biol 23:13–22

    Article  CAS  Google Scholar 

  • Kowalski R, Wojtczak M, Glogowski J, Ciereszko A (2003) Gelatinolytic and anti-trypsin activities in seminal plasma of common carp: relationship to blood, skin mucus and spermatozoa. Aquat Living Res 16:438–444

    Article  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head bacteriophage T4. Nature 277:680–685

    Article  Google Scholar 

  • Lowry OH, Rosenbrough NJ, Farr AR, Randall KJ (1951) Protein measurement with the folin phenol-reagent. J Biol Chem 193:265–275

    PubMed  CAS  Google Scholar 

  • Martins ML, Souza VN, Moraes JRE, Moraes FR, Costa AJ (1999) Comparative evaluation of the susceptibility of cultivated fishes to the natural infection with myxosporean parasites and tissue changes in the host. Braz J Biol 59:263–269

    Google Scholar 

Download references

Acknowledgements

We are grateful to Dr. Jayme Bastos (Biochemistry Department, UERJ-University of the State of Rio de Janeiro, Brazil) for the donation of tambacu from Morro Grande Farm. This work was supported by CNPq, Faperj and Oswaldo Cruz Foundation – Fiocruz.

Author information

Authors and Affiliations

  1. Laboratório de Química e Função de Proteínas e Peptídeos, Centro de Biociências e Biotecnologia/Universidade Estadual do Norte Fluminense, Av. Alberto Lamego, 2000, 28013-602, Campos dos Goytacazes, RJ, Brazil

    C. M. C. Salles

  2. Laboratório de Bioquímica de Proteínas e Peptídeos, DBBM/IOC/FIOCRUZ, Manguinhos, RJ, Brazil

    P. Gagliano, H. L. M. Guedes & S. Giovanni De-Simone

  3. Laboratório de Biotecnologia, CBB/UENF, Campos dos Goytacazes, RJ, Brazil

    J. B. Salles

  4. Laboratório de Bioquímica Toxicológica, IBRAG/UERJ, Rio de Janeiro, RJ, Brazil

    S. A. T. Leitão & V. P. F. Cassano

Authors
  1. C. M. C. Salles
    View author publications

    Search author on:PubMed Google Scholar

  2. P. Gagliano
    View author publications

    Search author on:PubMed Google Scholar

  3. S. A. T. Leitão
    View author publications

    Search author on:PubMed Google Scholar

  4. J. B. Salles
    View author publications

    Search author on:PubMed Google Scholar

  5. H. L. M. Guedes
    View author publications

    Search author on:PubMed Google Scholar

  6. V. P. F. Cassano
    View author publications

    Search author on:PubMed Google Scholar

  7. S. Giovanni De-Simone
    View author publications

    Search author on:PubMed Google Scholar

Corresponding author

Correspondence to C. M. C. Salles.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Salles, C.M.C., Gagliano, P., Leitão, S.A.T. et al. Identification and characterization of proteases from skin mucus of tambacu, a Neotropical hybrid fish. Fish Physiol Biochem 33, 173–179 (2007). https://doi.org/10.1007/s10695-007-9128-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10695-007-9128-7

Keywords