In order to investigate the possible link between PMCA (plasma-membrane Ca2+-ATPase) activity and D-glucose catabolism in insulin-producing cells, BRIN-BD11 cells were transfected with two isoforms of PMCA2. Transfection of insulin-producing BRIN-BD11 cells with PMCA2yb and PMCA2wb was documented by RT-PCR (reverse transcription-PCR), Western blot analysis, indirect immunofluorescence microscopy and 45Ca2+ uptake by microsomes. In the transfected cells, the overexpression of PMCA coincided with three major anomalies of D-glucose metabolism, namely a lower rate of D-[5-3H]glucose utilization prevailing at a low extracellular concentration of D-glucose (1.1 mM), a low ratio between D-[U-14C]oxidation and D-[5-3H]glucose utilization prevailing at a high extracellular glucose concentration (16.7 mM), and a high ratio between the net generation of 14C-labelled acidic metabolites and amino acids and that of 3H2O from D-[5-3H]glucose. These anomalies resulted in a decreased estimated rate of ATP generation (linked to the catabolism of the hexose) and a lowered ATP cell content, whether at low or high extracellular D-glucose concentrations. The net uptake of 45Ca2+ by intact cells was also decreased in the transfected cells, but to a greater extent than can apparently be attributed to the change in the ATP-generation rate. These findings document the relevance of PMCA activity to both D-glucose metabolism and Ca2+ handling in insulin-producing cells, with emphasis on the key role of both cytosolic and mitochondrial Ca2+ concentrations in the regulation of D-glucose catabolism. They also reveal that overexpression of PMCA leads, in insulin-producing cells, to an imbalance between ATP generation and consumption.
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September 11 2008
Effects of plasma membrane Ca2+-ATPase overexpression upon D-glucose metabolism in insulin-producing BRIN-BD11 cells
Adama Kamagate;
Adama Kamagate
*Laboratoire de Pharmacodynamie et de Thérapeutique, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium
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Abdullah Sener;
Abdullah Sener
†Laboratoire d'Hormonologie Expérimentale, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium
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Philippe Courtois;
Philippe Courtois
†Laboratoire d'Hormonologie Expérimentale, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium
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Willy J. Malaisse;
Willy J. Malaisse
1
†Laboratoire d'Hormonologie Expérimentale, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium
1To whom correspondence should be addressed (email [email protected]).
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André Herchuelz
André Herchuelz
*Laboratoire de Pharmacodynamie et de Thérapeutique, Université Libre de Bruxelles (ULB), 808 Route de Lennik, B-1070 Brussels, Belgium
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Publisher: Portland Press Ltd
Received:
May 27 2008
Revision Received:
July 18 2008
Accepted:
July 21 2008
Accepted Manuscript online:
July 21 2008
Online ISSN: 1573-4935
Print ISSN: 0144-8463
© The Authors Journal compilation © 2008 Biochemical Society
2008
Biosci Rep (2008) 28 (5): 251–258.
Article history
Received:
May 27 2008
Revision Received:
July 18 2008
Accepted:
July 21 2008
Accepted Manuscript online:
July 21 2008
Citation
Adama Kamagate, Abdullah Sener, Philippe Courtois, Willy J. Malaisse, André Herchuelz; Effects of plasma membrane Ca2+-ATPase overexpression upon D-glucose metabolism in insulin-producing BRIN-BD11 cells. Biosci Rep 1 October 2008; 28 (5): 251–258. doi: https://doi.org/10.1042/BSR20080042
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