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Characterization of an ATP diphosphohydrolase (EC 3.6.1.5) in synaptosomes from cerebral cortex of adult rats

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Abstract

Data from the literature have demonstrated that synaptosomal preparations from various sources can hydrolyze externally added ATP. Various authors characterized this activity as an ecto-ATPase. In the present report, we demonstrate that synaptosomal preparations obtained from the cerebral cortex of rats show ATPase activity that could not be dissociated from ADPase activity, suggesting that an ATP-diphosphohydrolase is involved in ATP and ADP hydrolysis. Furthermore, the ATP and ADP hydrolysis could not be attributed to associations of enzymes that could mimic an ATP-diphosphohydrolase because none of the following activities were detected in our assay conditions inorganic pyrophosphatase, adenylate kinase, or nonspecific phosphatases. A possible association between an ATPase and an ADPase was excluded on the basis of both the kinetics and much additional data on inhibitors, ion dependence, pH, etc. The present results demonstrate that in synaptosomal preparations from cerebral cortex an ATP-diphosphohydrolase is involved, at least in part, in ATP and ADP hydrolysis.

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Abbreviations

DCCD:

dicyclohexylcarboiimide

EDTA:

ethylenediaminetetraacetic acid

HEPES:

N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid

Pi:

inorganic phosphate

ATP:

diphosphohydrolase, Apyrase (EC 3.6.1.5)

ATPase:

ATP phosphohydrolase (EC 3.6.1.3) 5′-nucleotidase (EC 3.1.3.5) Hexokinase (EC 2.7.1.1) Glucose-6-phosphate dehydrogenase (EC 1.1.1.49) Adenylate kinase (EC 2.7.4.3) Inorganic pyrophosphatase (EC 3.6.1.1)

ATP:

pyrophosphohydrolase (EC 3.6.1.8)

LDH:

lactate dehydrogenase (EC 1.1.1.27)

SDH:

succinate dehydrogenase (EC 1.3.1.6)

ACHE:

acethylcholinesterase (EC 3.1.1.7)

G-6-Pase:

glucose-6-phosphatase (EC 3.1.3.9)

NADPH:

cytoehrome c oxidoreductase (NCR) (EC 1.6.2.4)

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Authors and Affiliations

  1. Departamento de Ciências Fisiológicas, Fundacão Universidade do Rio Grande, Rio Grande, RS, Brasil

    Ana M. O. Battastini

  2. Setor de Bioquímica-Centro de Ciências Naturais e Exatas, Universidade Federal de Santa Maria, Santa Maria, RS, Brasil

    João B. T. da Rocha

  3. Departamento de Bioquimica, Institute de Biociências, Universidade Federal do Rio Grande do Sul, rua Sarmento Leite, 500, 90050, Porto Alegre, RS, Brasil

    Claudia K. Barcellos, Renato D. Dias & João J. F. Sarkis

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Battastini, A.M.O., da Rocha, J.B.T., Barcellos, C.K. et al. Characterization of an ATP diphosphohydrolase (EC 3.6.1.5) in synaptosomes from cerebral cortex of adult rats. Neurochem Res 16, 1303–1310 (1991). https://doi.org/10.1007/BF00966661

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