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Salivary extracellular vesicles can modulate purinergic signalling in oral tissues by combined ectonucleoside triphosphate diphosphohydrolases and ecto-5′-nucleotidase activities

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Abstract

We reported previously that the rat submandibular gland is able to release nanovesicles capable to hydrolyse millimolar concentrations of ATP, ADP and AMP in vitro. Here, we show that rat saliva also contains nanovesicles with the ability to hydrolyse ATP. Our aim was to identify and characterize vesicular nucleotidases by using kinetic, immunological and in silico approaches. Nucleotidase activity in the absence or presence of specific inhibitors allowed us to assume the participation of NTPDase1, -2 and -3, together with ecto-5′-nucleotidase, confirmed using specific antibodies. At neutral pH, initial ATPase activity would be mostly due to NTPDase2, which was thereafter inactivated, leaving NTPDase1 and NTPDase3 to hydrolyse ATP and ADP with an efficacy ATPase/ADPase around 2. Ecto-5′nucleotidase would be mainly responsible for AMP hydrolysis and adenosine accumulation. We proposed a kinetic model for NTPDase2 as a tool to isolate and analyse the turnover of this enzyme in the presence of different ATP concentrations, including those expected in extracellular media. Our study characterizes the ectonucleotidases carried by extracellular vesicles which contribute to modulate ATP and adenosine concentrations in the oral cavity, essential players in purinergic signalling.

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Acknowledgements

This work was supported by the Universidad de Buenos Aires (UBACYT Grants 2002015020012BA and 20720150100006BA).

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Author notes

  1. Débora A. González and Martín M. Barbieri van Haaster have equally contributed to this work.

Authors and Affiliations

  1. Cátedra de Biofísica y Bioestadística, Facultad de Odontología, Universidad de Buenos Aires, M. T. de Alvear 2142, 1122, Buenos Aires, Argentina

    Débora A. González & Martín M. Barbieri van Haaster

  2. Cátedra de Farmacología, Facultad de Odontología, Universidad de Buenos Aires, M. T. de Alvear 2142, 1122, Buenos Aires, Argentina

    Emmanuel Quinteros Villarruel & Betina Orman

  3. Université de Paris, Integrated Biology of Red Blood Cell UMR_S1134, INSERM, 75015, Paris, France

    Claude Hattab & Mariano A. Ostuni

  4. Institut National de la Transfusion Sanguine (INTS), 6 Rue Alexandre Cabanel, 75015, Paris, France

    Claude Hattab & Mariano A. Ostuni

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  1. Débora A. González
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Correspondence to Débora A. González.

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González, D.A., Barbieri van Haaster, M.M., Quinteros Villarruel, E. et al. Salivary extracellular vesicles can modulate purinergic signalling in oral tissues by combined ectonucleoside triphosphate diphosphohydrolases and ecto-5′-nucleotidase activities. Mol Cell Biochem 463, 1–11 (2020). https://doi.org/10.1007/s11010-019-03624-6

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