Activating Compound | Comment | Organism | Structure |
---|---|---|---|
additional information | isozyme GAD65 is activated by phosphorylation | Homo sapiens | |
additional information | isozyme GAD65 is activated by phosphorylation | Rattus norvegicus |
Cloned (Comment) | Organism |
---|---|
gene GAD65, recombinant expression of GST-tagged wild-type and mutant isozymes in Escherichia coli strain DH5alpha or strain BL21 | Homo sapiens |
gene GAD67, recombinant expression of GST-tagged wild-type isozyme in Escherichia coli strain DH5alpha or strain BL21 | Homo sapiens |
Protein Variants | Comment | Organism |
---|---|---|
T95A | site-directed mutagenesis, the mutation T95A of the phosphorylation site abolishes the phosphorylation and its effects on enzyme activity | Homo sapiens |
T95E | site-directed mutagenesis, when the phosphorylation site T95 is mutated to glutamic acid, which mimics the phosphorylation status of hGAD65, the enzyme activity is greatly increased. An increase of GAD65 activity by 55% compared to the wild type hGAD65 is observed indicating that mutation of T95 to glutamic acid mimics the effect of phosphorylation | Homo sapiens |
Inhibitors | Comment | Organism | Structure |
---|---|---|---|
additional information | isozyme GAD67 is inhibited by phosphorylation | Homo sapiens |
Localization | Comment | Organism | GeneOntology No. | Textmining |
---|---|---|---|---|
membrane | - |
Homo sapiens | 16020 | - |
membrane | - |
Rattus norvegicus | 16020 | - |
soluble | - |
Homo sapiens | - |
- |
synaptosome | - |
Rattus norvegicus | - |
- |
Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | Homo sapiens | - |
4-aminobutanoate + CO2 | - |
? | |
L-glutamate | Rattus norvegicus | - |
4-aminobutanoate + CO2 | - |
? | |
L-glutamate | Rattus norvegicus Sprague Dawley | - |
4-aminobutanoate + CO2 | - |
? |
Organism | UniProt | Comment | Textmining |
---|---|---|---|
Homo sapiens | Q05329 | - |
- |
Homo sapiens | Q99259 | - |
- |
Rattus norvegicus | Q05683 | - |
- |
Rattus norvegicus Sprague Dawley | Q05683 | - |
- |
Posttranslational Modification | Comment | Organism |
---|---|---|
phosphoprotein | isozyme GAD65 is activated by phosphorylation at T95, identification of potential phosphorylation sites S3, S6, T95, and S417 on rGAD65 by tandem MS/MS spectrometry. The effect of phosphorylation on GAD65 can be reversed by treatment with protein phosphatases. Protein kinase C isoform epsilon is the protein kinase responsible for phosphorylation and regulation of GAD65 | Rattus norvegicus |
phosphoprotein | isozyme GAD65 is activated by phosphorylation at Thr95, while isozyme GAD67 is inhibited by phosphorylation. Gad65 is also phosphorylated at Ser417. The effect of phosphorylation on GAD65 and GAD67 can be reversed by treatment with protein phosphatases. Protein kinase A (PKA) and protein kinase C isoform epsilon are the protein kinases responsible for phosphorylation and regulation of GAD67 and GAD65, respectively | Homo sapiens |
phosphoprotein | isozyme GAD65 is activated by phosphorylation, while isozyme GAD67 is inhibited by phosphorylation. The effect of phosphorylation on GAD65 and GAD67 can be reversed by treatment with protein phosphatases. Protein kinase A (PKA) and protein kinase C isoform epsilon are the protein kinases responsible for phosphorylation and regulation of GAD67 and GAD65, respectively | Homo sapiens |
Purification (Comment) | Organism |
---|---|
preparation of native enzyme in rat brain synaptosomes, isolation of KCl-stimulated P2'-synaptosomes | Rattus norvegicus |
recombinant GST-tagged wild-type isozyme GAD65 from Escherichia coli strain DH5alpha by glutathione affinity chromatography and cleavage of the GST-tag by Factor Xa, followed by another step of glutathione affinity chromatography, or recombinant GST-tagged wild-type isozyme GAD65 from Escherichia coli strain BL21 by glutathione affinity chromatography and cleavage of the GST-tag by PreScission protease cleavage | Homo sapiens |
recombinant GST-tagged wild-type isozyme GAD67 from Escherichia coli strain DH5alpha by glutathione affinity chromatography and cleavage of the GST-tag by Factor Xa, followed by another step of glutathione affinity chromatography, or recombinant GST-tagged wild-type isozyme GAD67 from Escherichia coli strain BL21 by glutathione affinity chromatography and cleavage of the GST-tag by PreScission protease cleavage | Homo sapiens |
Source Tissue | Comment | Organism | Textmining |
---|---|---|---|
brain | - |
Homo sapiens | - |
brain | - |
Rattus norvegicus | - |
Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
---|---|---|---|---|---|---|
L-glutamate | - |
Homo sapiens | 4-aminobutanoate + CO2 | - |
? | |
L-glutamate | - |
Rattus norvegicus | 4-aminobutanoate + CO2 | - |
? | |
L-glutamate | - |
Rattus norvegicus Sprague Dawley | 4-aminobutanoate + CO2 | - |
? |
Synonyms | Comment | Organism |
---|---|---|
GAD1 | - |
Homo sapiens |
GAD2 | - |
Homo sapiens |
GAD2 | - |
Rattus norvegicus |
GAD65 | - |
Homo sapiens |
GAD65 | - |
Rattus norvegicus |
GAD67 | - |
Homo sapiens |
glutamate decarboxylase 65 | - |
Rattus norvegicus |
hGAD65 | - |
Homo sapiens |
hGAD65 | - |
Rattus norvegicus |
hGAD67 | - |
Homo sapiens |
L-glutamate decarboxylase 65 | - |
Homo sapiens |
L-glutamate decarboxylase 65 | - |
Rattus norvegicus |
L-glutamate decarboxylase 67 | - |
Homo sapiens |
Cofactor | Comment | Organism | Structure |
---|---|---|---|
pyridoxal 5'-phosphate | - |
Homo sapiens | |
pyridoxal 5'-phosphate | - |
Rattus norvegicus |
General Information | Comment | Organism |
---|---|---|
malfunction | phosphorylation site mutation T95A abolishes the phosphorylation and its effects on enzyme activity. When the phosphorylation site T95 is mutated to glutamic acid, which mimics the phosphorylation status of hGAD65, the enzyme activity is greatly increased. An increase of GAD65 activity by 55% compared to the wild type hGAD65 is observed indicating that mutation of T95 to glutamic acid mimics the effect of phosphorylation | Homo sapiens |
physiological function | isozyme GAD65 is activated by phosphorylation on Thr95. Protein kinase C isoform epsilon is the protein kinase responsible for phosphorylation and regulation of GAD65. Role of phosphorylation of GAD65 in regulation of GABA neurotransmission. Effect of neuronal stimulation on the level of membrane associated GAD (mGAD and GAD65) and soluble GAD (sGAD and GAD67), overview | Homo sapiens |
physiological function | isozyme GAD65 is activated by phosphorylation on Thr95. Protein kinase C isoform epsilon is the protein kinase responsible for phosphorylation and regulation of GAD65. Role of phosphorylation of GAD65 in regulation of GABA neurotransmission. Effect of neuronal stimulation on the level of membrane associated GAD (mGAD and GAD65) and soluble GAD (sGAD and GAD67), overview | Rattus norvegicus |
physiological function | isozyme GAD67 is inhibited by phosphorylation. Protein kinase A is the protein kinase responsible for phosphorylation and regulation of GAD67. Role of phosphorylation of GAD65 in regulation of GABA neurotransmission. Effect of neuronal stimulation on the level of membrane associated GAD (mGAD and GAD65) and soluble GAD (sGAD and GAD67), overview | Homo sapiens |