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Am J Physiol Endocrinol Metab 284: E399-E406, 2003; doi:10.1152/ajpendo.00259.2002
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Vol. 284, Issue 2, E399-E406, February 2003

Creatine transporter activity and content in the rat heart supplemented by and depleted of creatine

Ernest Boehm, Sharon Chan, Mina Monfared, Theo Wallimann, Kieran Clarke, and Stefan Neubauer

Wellcome Trust Centre for Human Genetics, University of Oxford, Oxford OX3 7BN; Department of Biochemistry, University of Oxford, Oxford OX1 3BN, United Kingdom; and Eidgenoessisch-technische Hochschule (ETH)-Zürich, Institute of Cell Biology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland

The intracellular creatine concentration is an important bioenergetic parameter in cardiac muscle. Although creatine uptake is known to be via a NaCl-dependent creatine transporter (CrT), its localization and regulation are poorly understood. We investigated CrT kinetics in isolated perfused hearts and, by using cardiomyocytes, measured CrT content at the plasma membrane or in total lysates. Rats were fed control diet or diet supplemented with creatine or the creatine analog beta -guanidinopropionic acid (beta -GPA). Creatine transport in control hearts followed saturation kinetics with a Km of 70 ± 13 mM and a Vmax of 3.7 ± 0.07 nmol · min-1 · g wet wt-1. Creatine supplementation significantly decreased the Vmax of the CrT (2.7 ± 0.17 nmol · min-1 · g wet wt-1). This was matched by an ~35% decrease in the plasma membrane CrT; the total CrT pool was unchanged. Rats fed beta -GPA exhibited a >80% decrease in tissue creatine and increase in beta -GPAtotal. The Vmax of the CrT was increased (6.0 ± 0.25 nmol · min-1 · g wet wt-1) and the Km decreased (39.8 ± 3.0 mM). The plasma membrane CrT increased about fivefold, whereas the total CrT pool remained unchanged. We conclude that, in heart, creatine transport is determined by the content of a plasma membrane isoform of the CrT but not by the total cellular CrT pool.

creatine transport(er); creatine feeding; creatine depletion; isolated perfused heart; cell-surface biotinylation


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