L(+)-Lactate binding to preparations of rat hepatocyte plasma membranes.
Incubation of rat hepatocyte plasma membranes with L-[14C]lactate resulted in the labeling of protein(s) of apparent molecular weight 40,000 when examined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The binding was saturable, irreversible, and inhibited by pyruvate, 2-oxoglutarate, and alpha-cyano-3-hydroxycinnamate, but not by D-lactate. It was markedly enhanced by L-alanine, but not D-alanine or beta-alanine. The binding protein(s) could be solubilized in cholic acid giving a single peak on gel filtration corresponding to a molecular weight of 26,000 and an isoelectric point of 5.1. This peak, when subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, ran in a position corresponding to an apparent molecular weight of 40,000. When membranes were treated with Triton X-100, lactate binding was retained by the Triton-insoluble fraction. The binding of L-[14C]lactate increased with incubation time, due apparently to the appearance of new binding sites and not to sequestration into vesicles. As many of the characteristics of lactate binding to rat hepatocyte plasma membranes were found to be similar to those of lactate entry into isolated hepatocytes, we speculate that the lactate-binding protein could represent part or whole of a plasma-membrane lactate transporter. Lactate-binding proteins of the same molecular weight were identified in the plasma membranes from rat erythrocytes, cardiac muscle, skeletal muscle, lung, and brain.
D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002458:Cell Fractionation; D002462:Cell Membrane / Q000378:metabolism / Q000648:ultrastructure; D007700:Kinetics; D007773:Lactates / Q000378:metabolism*; D019344:Lactic Acid; D008099:Liver / Q000378:metabolism*; D008297:Male; D008565:Membrane Proteins / Q000378:metabolism*; D027501:Monocarboxylic Acid Transporters; D051381:Rats; D011919:Rats, Inbred Strains; D012995:Solubility; D013237:Stereoisomerism
D002352:Carrier Proteins; D007773:Lactates; D008565:Membrane Proteins; D027501:Monocarboxylic Acid Transporters; D019344:Lactic Acid