title: Functional characterization of vesicular excitatory amino acid transport by human sialin.
abstract: Sialin, the protein coded by SLC17A5, is responsible for membrane potential (Δψ)-driven aspartate and glutamate transport into synaptic vesicles in addition to H+/sialic acid co-transport in lysosomes. Rodent sialin mutants harboring the mutations associated with Salla disease in humans did not transport aspartate and glutamate whereas H+/sialic acid co-transport activity was about one-third of the wild-type protein. In this study, we investigate the effects of various mutations on the transport activities of human sialin. Proteoliposomes containing purified heterologously expressed human sialin exhibited both Δψ-driven aspartate and glutamate transport activity and H+/sialic acid co-transport activity. Aspartate and glutamate transport was not detected in the R39C and K136E mutant forms of SLC17A5 protein associated with Salla disease, whereas H+/sialic acid co-transport activity corresponded to 30-50% of the recombinant wild-type protein. In contrast, SLC17A5 protein harboring the mutations associated with infantile sialic acid storage disease, H183R and Δ268SSLRN272 still showed normal levels of Δψ-driven aspartate and glutamate transport even though H+/sialic acid co-transport activity was absent. Human sialin carrying the G328E mutation that causes both phenotypes, and P334R and G378V mutations that cause infantile sialic acid storage disease showed no transport activity. These results support the idea that people suffering from Salla disease have been defective in aspartergic and glutamatergic neurotransmissions.
authors.affiliation: Advanced Science Research Center, Okayama University, Okayama, Japan. tmiyaji@pharm.okayama-u.ac.jp
authors.forename: Takaaki
authors.initials: T
authors.lastname: Miyaji
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Omote
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Moriyama
mesh_terms: D000818:Animals; D001224:Aspartic Acid / Q000378:metabolism; D016367:Baculoviridae / Q000378:metabolism; D001693:Biological Transport, Active / Q000502:physiology; D018076:DNA, Complementary / Q000096:biosynthesis / Q000235:genetics; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D008081:Liposomes / Q000737:chemistry; D008958:Models, Molecular; D008969:Molecular Sequence Data; D016296:Mutagenesis; D009154:Mutation / Q000502:physiology; D027361:Organic Anion Transporters / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D011994:Recombinant Proteins / Q000737:chemistry; D020133:Reverse Transcriptase Polymerase Chain Reaction; D029461:Sialic Acid Storage Disease / Q000235:genetics; D027981:Symporters / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D050492:Vesicular Biogenic Amine Transport Proteins / Q000378:metabolism*
chemical_list: D018076:DNA, Complementary; D008081:Liposomes; D027361:Organic Anion Transporters; D011994:Recombinant Proteins; D027981:Symporters; D050492:Vesicular Biogenic Amine Transport Proteins; C421224:sialic acid transport proteins; D001224:Aspartic Acid; D018698:Glutamic Acid