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Relationship between Ca2+-transport and ATP hydrolytic activities in guinea-pig pancreatic acinar plasma membranes

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Abstract

Partially purified plasma membrane fractions were prepared from guinea-pig pancreatic acini. These membrane preparations were found to contain an ATP-dependent Ca2+-transporter as well as a heterogenous ATP-hydrolytic activity. The Ca2+-transporter showed high affinity for Ca2+ (KCa 2+ = 0.04 ± 0.01 μM), an apparent requirement for Mg2+ and high substrate specificity. The major component of ATPase activity could be stimulated by either Ca2+ or Mg2+ but showed a low affinity for these cations. At low concentrations, Mg2+ appeared to inhibit the Ca2+-dependent ATPase activity expressed by these membranes. However, in the presence of high Mg2+ concentration (0.5–1 mM), a high affinity Ca2+-dependent ATPase activity was observed (KCa 2+ = 0.08 ± 0.02 μM). The hydrolytic activity showed little specificity towards ATP. Neither the Ca2+-transport nor high affinity Ca2+-ATPase activity were stimulated by calmodulin. The results demonstrate, in addition to a low affinity Ca2+ (or Mg+)-ATPase activity, the presence of both a high affinity Ca2+-pump and high affinity Ca2+-dependent ATPase. However, the high affinity Ca2+-ATPase activity does not appear to be the biochemical expression of the Ca2+-pump.

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Abbreviations

Ca2+-ATPase:

calcium-activated, magnesium-dependent adenosine triphosphatase

CaM:

calmodulin

CDTA:

trans-1,2-diaminocyclohexane-N,N,N′,N′-tetraacetate

EDTA:

ethylene-diaminetetraacetate

EGTA:

ethylene glycol bis(β-aminoethyl ether)-N,N,N′,N′-tetraacetate

NADPH:

reduced form of nicotinamide adenine dinucleotide phosphate

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Authors and Affiliations

  1. Division of Pharmacology and Toxicology, University of British Columbia, Vancouver, B. C., Canada

    Rajesh Mahey

  2. Division of Clinical Pharmacy, Faculty of Pharmaceutical Sciences, University of British Columbia, Vancouver, B. C., Canada

    Michael A. Bridges & Sidney Katz

  3. Departments of Paediatrics and Pathology, Faculty of Medicine, University of British Columbia, Vancouver, B. C., Canada

    Michael A. Bridges

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  1. Rajesh Mahey
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  2. Michael A. Bridges
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  3. Sidney Katz
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Mahey, R., Bridges, M.A. & Katz, S. Relationship between Ca2+-transport and ATP hydrolytic activities in guinea-pig pancreatic acinar plasma membranes. Mol Cell Biochem 105, 137–147 (1991). https://doi.org/10.1007/BF00227753

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  • DOI: https://doi.org/10.1007/BF00227753

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