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Na+,K+-ATPase Activity in the Posterior Gills of the Blue Crab, Callinectes ornatus (Decapoda, Brachyura): Modulation of ATP Hydrolysis by the Biogenic Amines Spermidine and Spermine

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Abstract

We investigated the effect of the exogenous polyamines spermine, spermidine and putrescine on modulation by ATP, K+, Na+, NH4 + and Mg2+ and on inhibition by ouabain of posterior gill microsomal Na+,K+-ATPase activity in the blue crab, Callinectes ornatus, acclimated to a dilute medium (21‰ salinity). This is the first kinetic demonstration of competition between spermine and spermidine for the cation sites of a crustacean Na+,K+-ATPase. Polyamine inhibition is enhanced at low cation concentrations: spermidine almost completely inhibited total ATPase activity, while spermine inhibition attained 58%; putrescine had a negligible effect on Na+,K+-ATPase activity. Spermine and spermidine affected both V and K for ATP hydrolysis but did not affect ouabain-insensitive ATPase activity. ATP hydrolysis in the absence of spermine and spermidine obeyed Michaelis–Menten behavior, in contrast to the cooperative kinetics seen for both polyamines. Modulation of V and K by K+, Na+, NH4 + and Mg2+ varied considerably in the presence of spermine and spermidine. These findings suggest that polyamine inhibition of Na+,K+-ATPase activity may be of physiological relevance to crustaceans that occupy habitats of variable salinity.

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Acknowledgments

This work constitutes part of a PhD thesis by D. P. G., financed by research grants from the Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP), the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), and the Instituto Nacional de Ciência e Tecnologia (INCT) Adapta/Fundação de Amparo à Pesquisa do Estado do Amazonas (FAPEAM, grant 573976/2008-2). F. A. L., C. F. L. F. and J. C. M. received research scholarships from CNPq. D. P. G. received a postdoctoral scholarship from FAPESP, and M. N. L. and J. L. F. received research scholarships from CNPq. All experiments conducted in this study complied with currently applicable state and federal laws. We thank Nilton Rosa Alves for technical assistance. This laboratory (F. A. L.) is integrated with the Amazon Shrimp Network (Rede de Camarão da Amazônia).

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Authors and Affiliations

  1. Departamento de Química, Faculdade de Filosofia, Ciências e Letras da Universidade de São Paulo, Ribeirão Preto, Avenida Bandeirantes, 3900, Ribeirão Preto, SP, 14040-901, Brazil

    Daniela P. Garçon & Francisco A. Leone

  2. Departamento de Bioquímica e Imunologia, Faculdade de Medicina de Ribeirão Preto da Universidade de São Paulo, Avenida Bandeirantes, 3900, Ribeirão Preto, SP, 14040-901, Brazil

    Malson N. Lucena & Juliana L. França

  3. Departamento de Biologia, Faculdade de Filosofia, Ciências e Letras da Universidade de São Paulo, Ribeirão Preto, SP, Brazil

    John C. McNamara

  4. Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, 21941-590, Brazil

    Carlos F. L. Fontes

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  1. Daniela P. Garçon
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  2. Malson N. Lucena
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  3. Juliana L. França
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  6. Francisco A. Leone
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Correspondence to Francisco A. Leone.

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Garçon, D.P., Lucena, M.N., França, J.L. et al. Na+,K+-ATPase Activity in the Posterior Gills of the Blue Crab, Callinectes ornatus (Decapoda, Brachyura): Modulation of ATP Hydrolysis by the Biogenic Amines Spermidine and Spermine. J Membrane Biol 244, 9–20 (2011). https://doi.org/10.1007/s00232-011-9391-5

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