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J Appl Physiol 97: 1059-1066, 2004. First published April 30, 2004; doi:10.1152/japplphysiol.00009.2004
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MCT1 confirmed in rat striated muscle mitochondria

C. Eric Butz,1 Grant B. McClelland,1,2 and George A. Brooks1

1Department of Integrative Biology, University of California, Berkeley, California 94720; and 2Department of Biology, McMaster University, Hamilton, Ontario L8S 4K1, Canada

Submitted 6 January 2004 ; accepted in final form 27 April 2004

We sought to test the hypothesis that monocarboxylate transporter isoform 1 (MCT1) is the inner mitochondrial membrane lactate/pyruvate transporter, and, as such, contributes to functioning of the intracellular lactate shuttle. However, presence of a mammalian mitochondrially localized MCT1 (mMCT1) has been contested. We sought to confirm by Western blotting the mitochondrial localization of MCT1 in rat cardiac, soleus, and extensor digitorum longus muscles utilizing three different cell fractionation methods and three different antibodies. We performed Western blotting using antibodies to cell membrane glucose transporter isoform GLUT1, inner mitochondrial constituent cytochrome oxidase, the monocarboxylate transporter protein chaperone CD147, as well as custom and commercially available MCT1 antibodies. Western blots demonstrated similar results with each MCT1 antibody and two of three methods of fractionation. MCT1 was found in the mitochondria, as well as in the sarcolemmal membrane and whole muscle homogenates. Probing with GLUT1 and CD147 demonstrated that mitochondrial fractions were not contaminated with sarcolemmal remnants. Probing with cytochrome oxidase showed mitochondrial localization of MCT1. Comparison of these results to the findings of others indicates that the most likely source of discrepancy is the cell fractionation procedure utilized.

lactate; lactate shuttle; monocarboxylate transporter; monocarboxylate transporter isoform 1


Address for reprint requests and other correspondence: G. A. Brooks, Exercise Physiology Laboratory, Dept. of Integrative Biology, 5101 Valley Life Science Bldg., University of California, Berkeley, CA 94720-3140 (E-mail: gbrooks{at}socrates.berkeley.edu).




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