title: Transport activity of the high-affinity monocarboxylate transporter MCT2 is enhanced by extracellular carbonic anhydrase IV but not by intracellular carbonic anhydrase II.
abstract: The ubiquitous enzyme carbonic anhydrase isoform II (CAII) has been shown to enhance transport activity of the proton-coupled monocarboxylate transporters MCT1 and MCT4 in a non-catalytic manner. In this study, we investigated the role of cytosolic CAII and of the extracellular, membrane-bound CA isoform IV (CAIV) on the lactate transport activity of the high-affinity monocarboxylate transporter MCT2, heterologously expressed in Xenopus oocytes. In contrast to MCT1 and MCT4, transport activity of MCT2 was not altered by CAII. However, coexpression of CAIV with MCT2 resulted in a significant increase in MCT2 transport activity when the transporter was coexpressed with its associated ancillary protein GP70 (embigin). The CAIV-mediated augmentation of MCT2 activity was independent of the catalytic activity of the enzyme, as application of the CA-inhibitor ethoxyzolamide or coexpressing the catalytically inactive mutant CAIV-V165Y did not suppress CAIV-mediated augmentation of MCT2 transport activity. Furthermore, exchange of His-88, mediating an intramolecular H(+)-shuttle in CAIV, to alanine resulted only in a slight decrease in CAIV-mediated augmentation of MCT2 activity. The data suggest that extracellular membrane-bound CAIV, but not cytosolic CAII, augments transport activity of MCT2 in a non-catalytic manner, possibly by facilitating a proton pathway other than His-88.
authors.affiliation: Abteilung für Allgemeine Zoologie, Fachbereich Biologie, Technische Universität Kaiserslautern, P. O. Box 3049, 67653 Kaiserslautern, Germany.
authors.forename: Michael
authors.initials: M
authors.lastname: Klier
authors.forename: Christina
authors.initials: C
authors.lastname: Schüler
authors.forename: Andrew P
authors.initials: AP
authors.lastname: Halestrap
authors.forename: William S
authors.initials: WS
authors.lastname: Sly
authors.forename: Joachim W
authors.initials: JW
authors.lastname: Deitmer
authors.forename: Holger M
authors.initials: HM
authors.lastname: Becker
mesh_terms: D000818:Animals; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism; D024402:Carbonic Anhydrase II / Q000378:metabolism*; D030741:Carbonic Anhydrase IV / Q000378:metabolism*; D017931:DNA Primers; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007150:Immunohistochemistry; D027501:Monocarboxylic Acid Transporters / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D021381:Protein Transport; D014981:Xenopus
chemical_list: D017931:DNA Primers; D027501:Monocarboxylic Acid Transporters; C490427:SLC16A7 protein, human; D001710:Biotin; D024402:Carbonic Anhydrase II; D030741:Carbonic Anhydrase IV