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Elustondo 2013 J Biol Chem

From Bioblast
Publications in the MiPMap
Elustondo PA, White AE, Hughes ME, Brebner K, Pavlov E, Kane DA (2013) Physical and functional association of lactate dehydrogenase (LDH) with skeletal muscle mitochondria. J Biol Chem 288:25309-17.

ยป PMID: 23873936 Open Access

Elustondo PA, White AE, Hughes ME, Brebner K, Pavlov E, Kane DA (2013) J Biol Chem

Abstract: The intracellular lactate shuttle hypothesis posits that lactate generated in the cytosol is oxidized by mitochondrial lactate dehydrogenase (LDH) of the same cell. To examine whether skeletal muscle mitochondria oxidize lactate, mitochondrial respiratory oxygen flux (JO2) was measured during the sequential addition of various substrates and cofactors onto permeabilized rat gastrocnemius muscle fibers, as well as isolated mitochondrial subpopulations. Addition of lactate did not alter JO2. However, subsequent addition of NAD(+) significantly increased JO2, and was abolished by the inhibitor of mitochondrial pyruvate transport, ฮฑ-cyano-4-hydroxycinnamate. In experiments with isolated subsarcolemmal and intermyofibrillar mitochondrial subpopulations, only subsarcolemmal exhibited NAD(+)-dependent lactate oxidation. To further investigate the details of the physical association of LDH with mitochondria in muscle, immunofluorescence/confocal microscopy and immunoblotting approaches were used. LDH clearly colocalized with mitochondria in intact, as well as permeabilized fibers. LDH is likely localized inside the outer mitochondrial membrane, but not in the mitochondrial matrix. Collectively, these results suggest that extra-matrix LDH is strategically positioned within skeletal muscle fibers to functionally interact with mitochondria. โ€ข Keywords: Immunohistochemistry, Lactate Shuttle, Lactic Acid, Mitochondrial Metabolism, NAD, Pyruvate, Respiration, Respiratory Chain, Respirometry, Skeletal Muscle Metabolism

โ€ข O2k-Network Lab: CA Antigonish Kane DA


Organism: Rat  Tissue;cell: Skeletal muscle  Preparation: Permeabilized cells, Isolated mitochondria 

HRR: Oxygraph-2k