title: The inhibition of monocarboxylate transporter 2 (MCT2) by AR-C155858 is modulated by the associated ancillary protein.
abstract: In mammalian cells, MCTs (monocarboxylate transporters) require association with an ancillary protein to enable plasma membrane expression of the active transporter. Basigin is the preferred binding partner for MCT1, MCT3 and MCT4, and embigin for MCT2. In rat and rabbit erythrocytes, MCT1 is associated with embigin and basigin respectively, but its sensitivity to inhibition by AR-C155858 was found to be identical. Using RT (reverse transcription)-PCR, we have shown that Xenopus laevis oocytes contain endogenous basigin, but not embigin. Co-expression of exogenous embigin was without effect on either the expression of MCT1 or its inhibition by AR-C155858. In contrast, expression of active MCT2 at the plasma membrane of oocytes was significantly enhanced by co-expression of exogenous embigin. This additional transport activity was insensitive to inhibition by AR-C155858 unlike that by MCT2 expressed with endogenous basigin that was potently inhibited by AR-C155858. Chimaeras and C-terminal truncations of MCT1 and MCT2 were also expressed in oocytes in the presence and absence of exogenous embigin. L-Lactate Km values for these constructs were determined and revealed that the TM (transmembrane) domains of an MCT, most probably TM7-TM12, but not the C-terminus, are the major determinants of L-lactate affinity, whereas the associated ancillary protein has little or no effect. Inhibitor titrations of lactate transport by these constructs indicated that embigin modulates MCT2 sensitivity to AR-C155858 through interactions with both the intracellular C-terminus and TMs 3 and 6 of MCT2. The C-terminus of MCT2 was found to be essential for its expression with endogenous basigin.
authors.affiliation: Department of Biochemistry, School of Medical Sciences, University of Bristol, University Walk, Bristol, UK.
authors.forename: Matthew J
authors.initials: MJ
authors.lastname: Ovens
authors.forename: Christine
authors.initials: C
authors.lastname: Manoharan
authors.forename: Marieangela C
authors.initials: MC
authors.lastname: Wilson
authors.forename: Clarey M
authors.initials: CM
authors.lastname: Murray
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Halestrap
mesh_terms: D000818:Animals; D051926:Basigin / Q000378:metabolism; D004912:Erythrocytes / Q000187:drug effects / Q000378:metabolism; D006023:Glycoproteins / Q000378:metabolism*; D007700:Kinetics; D019344:Lactic Acid / Q000378:metabolism; D008565:Membrane Proteins; D027501:Monocarboxylic Acid Transporters / Q000037:antagonists & inhibitors* / Q000737:chemistry; D050505:Mutant Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry; D009865:Oocytes / Q000166:cytology / Q000187:drug effects; D011817:Rabbits; D051381:Rats; D011994:Recombinant Proteins / Q000378:metabolism; D027981:Symporters / Q000037:antagonists & inhibitors / Q000737:chemistry; D013876:Thiophenes / Q000494:pharmacology*; D014498:Uracil / Q000031:analogs & derivatives* / Q000494:pharmacology; D029867:Xenopus Proteins / Q000378:metabolism; D014982:Xenopus laevis / Q000378:metabolism; D000099233:Monocarboxylate Transport Protein 1
chemical_list: D051926:Basigin; D006023:Glycoproteins; D019344:Lactic Acid; D008565:Membrane Proteins; D027501:Monocarboxylic Acid Transporters; D050505:Mutant Proteins; D011994:Recombinant Proteins; D027981:Symporters; D013876:Thiophenes; D014498:Uracil; D029867:Xenopus Proteins; D000099233:Monocarboxylate Transport Protein 1; C546977:AR C155858; C504567:Emb protein, rat; C091633:Slc16a7 protein, rat