Cold-shock-stimulated γ-aminobutyric acid synthesis is mediated by an increase in cytosolic Ca2+, not by an increase in cytosolic H+
作者:
Ewa Cholewa,
Alan W. Bown,
Andrzej J. Cholewinski,
Barry J. Shelp,
Wayne A. Snedden,
期刊:
Canadian Journal of Botany
(NRC Available online 1997)
卷期:
Volume 75,
issue 3
页码: 375-382
ISSN:0008-4026
年代: 1997
DOI:10.1139/b97-040
出版商: NRC Research Press
数据来源: NRC
摘要:
Synthesis of the nonprotein amino acid γ-aminobutyric acid is stimulated within minutes by diverse environmental factors. Synthesis (L-Glu + H+– γ-aminobutyric acid + CO2) is catalysed byL-Glu decarboxylase, a cytosolic enzyme having an acidic pH optimum. This study uses isolatedAsparagus sprengeri(Regel) mesophyll cells to investigate the possible role of Ca2+in stimulated γ-aminobutyric acid synthesis. Abrupt cold shock (20 °C to 1 °C) stimulated γ-aminobutyric acid levels from 2.7 to 5.6 nmol/106cells within 15 min. This 100% increase was reduced to 28% in the presence of the Ca2+channel blocker lanthanum, and was significantly reduced by incubation with 1 mM of the calmodulin antagonistN-(6-aminohexyl)-5-chloro-1-naphthalene-sulfonamide. Incubation at 20 °C with 25 μM calcimycin, a Ca2+ionophore, increased levels by 61% within 15 min. A fluorescent Ca2+indicator demonstrated that cytosolic Ca2+increased within 2 s of cold shock, followed by a return to initial levels within 25 s. In contrast, comparable experiments indicate a rapid and prolonged decrease in cytosolic H+.L-Glu decarboxylase isolated from asparagus cladophylls was stimulated 100% by addition of 500 μM Ca2+and 200 nM calmodulin. This activity was reduced to control values by the calmodulin antagonist. Collectively, the data indicate that cold shock initiates a signal transduction pathway in which increased cytosolic Ca2+stimulates calmodulin-dependentL-Glu decarboxylase activity and γ-aminobutyric acid synthesis. This mechanism appears independent of increased H+.Key words: cold shock, GABA, Ca2+, H+.
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