Purification of the lysosomal sialic acid transporter. Functional characteristics of a monocarboxylate transporter
Sialic acid and glucuronic acid are monocarboxylated monosaccharides, which are normally present in sugar side chains of glycoproteins, glycolipids, and glycosaminoglycans. After degradation of these compounds in lysosomes, the free monosaccharides are released from the lysosome by a specific membrane transport system. This transport system is deficient in the human hereditary lysosomal sialic acid storage diseases (Salla disease and infantile sialic acid storage disease, OMIM 269920). The lysosomal sialic acid transporter from rat liver has now been purified to apparent homogeneity in a reconstitutively active form by a combination of hydroxyapatite, lectin, and ion exchange chromatography. A 57-kDa protein correlated with transport activity. The transporter recognized structurally different types of acidic monosaccharides, like sialic acid, glucuronic acid, and iduronic acid. Transport of glucuronic acid was inhibited by a number of aliphatic monocarboxylates (i.e. lactate, pyruvate, and valproate), substituted monocarboxylates, and several dicarboxylates. cis-Inhibition, trans-stimulation, and competitive inhibition experiments with radiolabeled glucuronic acid as well as radiolabeled L-lactate demonstrated that L-lactate is transported by the lysosomal sialic acid transporter. L-Lactate transport was proton gradient-dependent, saturable with a Km of 0.4 mM, and mediated by a single mechanism. These data show striking biochemical and structural similarities of the lysosomal sialic acid transporter with the known monocarboxylate transporters of the plasma membrane (MCT1, MCT2, MCT3, and Mev).
|Keywords||*Organic Anion Transporters, *Plant Lectins, *Symporters, Animals, Biological Transport, Carboxylic Acids/metabolism, Cell Fractionation, Chromatography, Chromatography, Affinity, Chromatography, Ion Exchange, Durapatite, Humans, Intracellular Membranes/metabolism/ultrastructure, Lectins, Liver/*metabolism, Lysosomal Storage Diseases/genetics, Lysosomes/*metabolism/ultrastructure, Membrane Transport Proteins/genetics/*isolation & purification/*metabolism, Monosaccharides/metabolism, Rats, Rats, Wistar, Research Support, Non-U.S. Gov't, Substrate Specificity|
Havelaar, A.C., Mancini, G.M.S., Beerens, C.E.M.T., Souren, R.M., & Verheijen, F.W.. (1998). Purification of the lysosomal sialic acid transporter. Functional characteristics of a monocarboxylate transporter. Journal of Biological Chemistry. Retrieved from http://hdl.handle.net/1765/8978