Inhibition of L-glutamic acid decarboxylase by cycloglutamates

Lp, Sastchenko; Severin, E. S.; De, Metzler; Rm, Khomutov

doi:10.1021/bi00802a009

Cited by 10 publications

(2 citation statements)

References 7 publications

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“…-Methyl-DL-glutamic acid has been shown previously to be slowly decarboxylated by the enzyme, and it also inactivates the enzyme by transaminating the pyridoxal phosphate (Sukhareva and Torchinsky, 1966;Huntley and Metzler, 1968). It has been suggested that glutamate decarboxylase undergoes a conformational change upon interaction with glutamic acid (Sastchenko et at. 1971).…”

Section: Discussionmentioning

confidence: 99%

Glutamate decarboxylase. Substrate specificity and inhibition by carboxylic acids

Fonda¹

1972

Biochemistry

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Section: Discussionmentioning

confidence: 99%

Glutamate decarboxylase. Substrate specificity and inhibition by carboxylic acids

Fonda¹

1972

Biochemistry

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“…4 ) Glutamate decarboxylase activity was determined in a Warburg apparatus. 5 ) Phosphorylase activity was assayed according to Cori et al,6) and glutamine synthetase was assayed by the determination of production of y-glutamylhydroxamate. 7 ) Glutamate dehydrogenase was assayed by the method of Fahien et al,8) and y-aminobutylate aminotransferase was assayed in a system coupled with succinic semialdehyde dehydrogenase.…”

Section: Methodsmentioning

confidence: 99%

Isolation and Some Properties of an Aspartate Aminotransferase Inhibitor, Gostatin

Nishino

Murao

1983

Agricultural and Biological Chemistry

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A novel aspartate aminotransferase inhibitor named gostatin was isolated as crystals from the culture filtrate of Streptomyces sumanensis . The structure has been established to be 5-amino-2-carboxy-4-oxo-l ,4,5,6-tetrahydropyridine-3-acetic acid by X-ray crystallographic analysis (K. Ibata et al., manuscript in preparation). The results of chemical and instrumental analyses of the inhibitor are described in this paper.Gostatin showed time-dependent inhibitory action against aspartate aminotransferases and alanine aminotransferase. Both pyridoxal phosphate-and pyridoxamine phosphate-linked enzymes were sensitive to the inhibitor. 1961A novel cyclic amino acid named gostatin, produced by Streptomy'ces sumanensis NK-23, shows a strong inhibitory activity against aspartate aminotransferases (Be 2.6.1.1, GOT)* of various origins.! ,2) This inhibitor was isolated as crystals and the structure was determined to be 5-amino-2-carboxy-4-oxo-l ,4,5,6-tetrahydropyridine-3-acetic acid ( Fig. 1) by Xray crystallographic analysis (K. Ibata et al., manuscript in preparation).In this paper, the isolation and some chemical and biochemical properties of gostatin are reported. MATERIALS AND METHODSMicroorganism and culture conditions. Streptomyces sumanensis NK-23, which was isolated from soil by the authors,1,2) was used in this experiment. The strain was cultured under the optimum conditions for the production of gostatin in a Marubishi MSJ-U2 jar fermentor. 2 ) The culture filtrate at 114 hr cultivation was used for the preparation of the inhibitor.Assay ofgostatin activity. Gostatin activity was assayed by the method described in the previous paper. 2 ) Isolation ofgostatin. The isolation procedure reported in the previous paper was modified. 1 ) The culture filtrate (35 liters) was adjusted to pH 4.0 with formic acid, and activated charcoal (2%, w/v) was added. After the mixture stood for 30 min, the non~adsorbingfraction was collected by filtration .. This solution was applied onto a column (10x35cm) of Dowex lx4 (formate type, 100",200 mesh) previously equilibrated with 50 mM ammonium formate-formic acid buffer (pH 4.0). Gostatin was weakly adsorbed to the column and eluted with 50 mM ammonium formate-formic acid buffer (pH 4.0). Active fractions thus obtained (36 liters) were then applied onto a column (10 x 35cm) of Dowex 1 x4 (OH type, 100",200 mesh) and eluted with 0.5 M NH 4 CI solution. The fractions containing gostatin were pooled (14 liters), evaporated in vacuo to dryness and dissolved in a minimum volume of water (1150 ml). After the solution was adjusted to pH 4.0 with 6N HCI, methanol was added at a final concentration of 50% (v/v) and the precipitate was removed by filtration. Gostatin was precipitated from the supernatant by adding ethanol at a final concentration of 60% and allowing the mixture to stand overnight. The precipitates were collected by centrifugation (10,000 x g, 10 min) and dissolved in 170 ml of water. Gostatin was crystallized from this solution by adjusting the pH (pH 4.0) and adding methanol...

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Probing the Active Sites of Enzymes with Conformationally Restricted Substrate Analogs

Kenyon

Fee

1973

Progress in Physical Organic Chemistry

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Inhibition of L-glutamic acid decarboxylase by cycloglutamates

Cited by 10 publications

References 7 publications

Glutamate decarboxylase. Substrate specificity and inhibition by carboxylic acids

Glutamate decarboxylase. Substrate specificity and inhibition by carboxylic acids

Isolation and Some Properties of an Aspartate Aminotransferase Inhibitor, Gostatin

Probing the Active Sites of Enzymes with Conformationally Restricted Substrate Analogs

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