title: Synaptic vesicular glutamate uptake: modulation by a synaptosomal cytosolic factor.
abstract: We have demonstrated previously that L-glutamate is taken up into isolated synaptic vesicles in an ATP-dependent manner, supporting the neurotransmitter role of this acidic amino acid. We now report that a nerve terminal cytosolic factor inhibits the ATP-dependent vesicular uptake of glutamate in a dose-dependent manner. This factor appears to be a protein with a molecular weight greater than 100,000, as estimated by size exclusion chromatography, and is precipitated by ammonium sulfate (40% saturation). The inhibitory factor is inactivated by heating to 100 degrees C. Proteolytic digestion of the ammonium sulfate fraction by trypsin or chymotrypsin did not reduce, but rather increased slightly, the inhibition of glutamate uptake. Unlike the native factor, the digest retained inhibitory activity after heating, suggesting that proteolytic digestion may generate active fragments. The inhibition of ATP-dependent vesicular glutamate uptake is not species-specific, as the factor obtained from both rat and bovine brains produced an equal degree of inhibition of glutamate uptake into vesicles of each species. These observations raise the possibility that vesicular uptake of glutamate may be regulated by an endogenous factor in vivo.
authors.affiliation: Department of Pharmacology, University of Michigan, Ann Arbor 48109.
authors.forename: A T
authors.initials: AT
authors.lastname: Lobur
authors.forename: P E
authors.initials: PE
authors.lastname: Kish
authors.forename: T
authors.initials: T
authors.lastname: Ueda
mesh_terms: D000818:Animals; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D003600:Cytosol / Q000378:metabolism*; D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates / Q000378:metabolism*; D018698:Glutamic Acid; D009419:Nerve Tissue Proteins / Q000502:physiology*; D009994:Osmolar Concentration; D010447:Peptide Hydrolases / Q000494:pharmacology; D051381:Rats; D013045:Species Specificity; D013572:Synaptic Vesicles / Q000378:metabolism*; D013574:Synaptosomes / Q000378:metabolism*
chemical_list: D018691:Excitatory Amino Acid Antagonists; D005971:Glutamates; D009419:Nerve Tissue Proteins; D018698:Glutamic Acid; D010447:Peptide Hydrolases