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Characteristics of Cl−-dependentl-[35S]cysteic acid transport into rat brain synaptic membrane vesicles

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Abstract

Uptake ofl-[35S]cysteic acid (L-CA) in rat synaptic membrane vesicles was investigated. Preincubation with either 10 mMl-glutamic acid (L-Glu), 25 mM L-CA, 10 mMdl-homocysteic acid, or 25 mMdl-2-amino-4-phosphonobutyrate on membrane vesicles enhanced L-[35S]CA and L-[3H]Glu uptake. Na+ (5 mM) and omission of Cl− from the assay medium decreased L-[35S]CA uptake into both 10 mM L-Glu-loaded and non-loaded membrane vesicles. The anion transport blockers, 4-acetamide-4′-isothiocyano-2,2′-disulfonic acid stibene (SITS) and 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene (DIDS), inhibited L-[35S]CA uptake in a dose-dependent manner. The maximal uptake rate for L-[35S]CA was decreased by 50 μM SITS, while the apparent Km value of L-CA was not changed. SITS increased the EC50 value of Cl− for L-[35S]CA uptake from 5 mM to 10 mM with reduction of the maximal effect. These results suggested that L-[35S]CA uptake into synaptic membrane vesicles was mediated by a SITS-sensitive hetero-exchange transport with non-labeled substrates.

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Abbreviations

SITS:

4-Acetamide-4′-isothiocyano-2,2′-disulfonic acid stilbene

DIDS:

4,4′-Diisothiocyano-2,2′-disulfonic acid stilbene

CA:

Cysteic acid

APB:

2-Amino-4-phosphonobutyrate

CSA:

Cysteine sulfinic acid

EGTA:

Ethyleneglycol bis(aminoethylether) tetraacetate

GABA:

γ-Aminobutyric acid

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Authors and Affiliations

  1. Department of Pharmacology, Faculty of Pharmaceutical Sciences, Osaka University, 1-6 Yamada-Oka, 565, Suita, Osaka, Japan

    Yutaka Koyama, Akemichi Baba & Heitaroh Iwata

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  1. Yutaka Koyama
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  2. Akemichi Baba
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  3. Heitaroh Iwata
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Koyama, Y., Baba, A. & Iwata, H. Characteristics of Cl−-dependentl-[35S]cysteic acid transport into rat brain synaptic membrane vesicles. Neurochem Res 15, 1153–1158 (1990). https://doi.org/10.1007/BF01208574

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