Yasuo Suda

Effect of aspartame on N-methyl-D-aspartate-sensitive L-[3H]glutamate binding sites in rat brain synaptic membranes.

Aspartame (L-aspartyl-L-phenylalanine methyl ester), an artificial low-calorie sweetener, was shown to dose-dependently inhibit L-[3H]glutamate binding to its N-methyl-D-aspartate-specific receptors. L-Aspartic acid, a major endogenous metabolite of aspartame, inhibited the binding more stronger than aspartame, while the other metabolites, L-phenylalanine and methanol, had no effect at the same concentration. Aspartame caused a significant change in the affinities of L-[3H]glutamate binding without altering the Vmax values of the binding, suggesting the inhibition is competitive. These in vitro findings suggested that aspartame may act directly on the N-methyl-D-aspartate-sensitive glutamate recognition sites in the brain synaptic membranes.

Molecular action mechanism of spider toxin on glutamate receptor: role of 2,4-dihydroxyphenylacetic acid in toxin molecule

Joro spider toxin (JSTX) isolated from Nephila clavata was shown to inhibit L-glutamate binding to rat brain synaptic membranes in a dose-dependent manner. 2,4-Dihydroxyphenylacetic acid (2,4-DHPA), a common moiety of spider toxins, also inhibited specifically L-glutamate binding at a concentration similar to that of the toxin. The binding activity inhibited by 2,4-DHPA or JSTX was recoverable on addition of ferric compound. These results suggest that 2,4-DHPA is a functional moiety in the toxin molecule and the biological action of spider toxin is explained by direct interaction with an Fe-S center which is known to play an important role for the glutamate binding.

A spider toxin (JSTX) inhibits l-glutamate uptake by rat brain synaptosomes ☆

Joro spider toxin (JSTX), a specific blocker of glutamate receptors, was found to exert a prominent suppressive action on the Na+-dependent binding of L-glutamate to synaptic membranes and on glutamate uptake by synaptosomes in a dose-dependent manner. In contrast, the synthesized 2,4-dihydroxyphenylacetylasparagine (2,4-DHPA-ASN), a common moiety of spider toxins, which has been shown to exhibit almost the same activity as intact JSTX with respect to the inhibition of Na+-independent glutamate binding to its synaptic membrane receptors, shows lower potency in inhibiting Na+-dependent binding and uptake of L-glutamate. From these findings, it is clear that JSTX has the ability to inhibit not only L-glutamate binding to its synaptic membrane receptors but also L-glutamate uptake by synaptosomes, and that polyamines linked to 2,4-DHPA-ASN in the molecule of spider toxins may participate in the inhibition of L-glutamate uptake.

Inhibitory effect of 2,4-dihydroxyphenylacetylasparagine, a common moiety of spider toxin, on glutamate binding to rat brain synaptic membranes

2,4-Dihydroxyphenylacetylasparagine (2,4-DHPA-ASN), a common moiety of molecules of spider toxins, was shown to inhibit L-[3H]glutamic acid binding to rat brain synaptic membranes in a dose-dependent manner. The inhibitory effect of 2,4-DHPA-ASN was almost the same as that of intact spider toxin isolated from Nephila clavata, but significantly higher than that of 2,4-dihydroxyphenylacetic acid (2,4-DHPA). In addition, neither 2,4-dihydroxybenzoic acid nor the isomers of 2,4-DHPA suppressed the glutamate binding. These results suggested that 2,4-DHPA might be the functional part and asparagine in the molecules of spider toxins seemed to cause increasing affinity toward the recognition site of glutamate binding.

Selective Labeling of Bioactive Polysaccharides with a New Fluorescent Reagent

The selective labeling of polysaccharides by the hydroxyl groups and without loosing biological activity is described. The new reagent is a deriv ative of fluoresceinamine and should be useful for the appropriate labeling of several types of biological active materials containing both amino and hydroxyl functional groups.

The Synthesis and Properties of Block-Copoly(amino acid)s with Folded Structures: A Primitive Enzyme Model

As a model of a primitive enzyme block-copoly(amino acid)s con taining L-serine and L-aspartic acid as the repeat unit were prepared. Spectro scopic measurements indicate that some block-copoly(amino acid)s, especially ABAB and ABABA types, do form folded conformations. Hydrolysis reactions of p-nitrophenyl p-guanidinobenzoate were accelerated in the presence of ABAB and ABABA type block-copoly(amino acid)s compared with a random copoly (amino acid). From these results we speculate that primitive enzymes were born by assembling block-copoly(amino acid)s.