NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1
The Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 contains two distinct glutamate dehydrogenases (GDH), each specific for either NADP + or NAD + . This feature is quite unusual in bacteria, which generally have a single GDH. NADP + ‐dependent GDH has been purified to homogeneity and the...
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crwiley:10.1046/j.1432-1327.2000.00972.x 2024-09-15T17:46:19+00:00 NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 Characterization, protein and DNA sequence, and relationship to other glutamate dehydrogenases Di Fraia, Raffaela Wilquet, Valerie Ciardiello, M. Antonietta Carratore, Vito Antignani, Antonella Camardella, Laura Glansdorff, Nicolas di Prisco, Guido 2000 http://dx.doi.org/10.1046/j.1432-1327.2000.00972.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1432-1327.2000.00972.x https://febs.onlinelibrary.wiley.com/doi/pdf/10.1046/j.1432-1327.2000.00972.x en eng Wiley http://onlinelibrary.wiley.com/termsAndConditions#vor European Journal of Biochemistry volume 267, issue 1, page 121-131 ISSN 0014-2956 1432-1033 journal-article 2000 crwiley https://doi.org/10.1046/j.1432-1327.2000.00972.x 2024-08-06T04:12:41Z The Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 contains two distinct glutamate dehydrogenases (GDH), each specific for either NADP + or NAD + . This feature is quite unusual in bacteria, which generally have a single GDH. NADP + ‐dependent GDH has been purified to homogeneity and the gene encoding GDH has been cloned and expressed. The enzyme has a hexameric structure. The amino acid sequence determined by peptide and gene analyses comprises 447 residues, yielding a protein with a molecular mass of 49 285 Da. The sequence shows homology with hexameric GDHs, with identity levels of 52% and 49% with Escherichia coli and Clostridium symbiosum GDH, respectively. The coenzyme‐binding fingerprint motif GXGXXG/A (common to all GDHs) has Ser at the last position in this enzyme. The overall hydrophilic character is increased and a five‐residue insertion in a loop between two α‐helices may contribute to the increase in protein flexibility. Psychrobacter sp. TAD1 GDH apparent temperature optimum is shifted towards low temperatures, whereas irreversible heat inactivation occurs at temperatures similar to those of E. coli GDH. The catalytic efficiency in the temperature range 10–30 °C is similar or lower than that of E. coli GDH. Unlike E. coli GDH the enzyme exhibits marked positive cooperativity towards 2‐oxoglutarate and NADPH. This feature is generally absent in prokaryotic GDHs. These observations suggest a regulatory role for this GDH, the most crucial feature being the structural/functional properties required for fine regulation of activity, rather than the high catalytic efficiency and thermolability encountered in several cold‐active enzymes. Article in Journal/Newspaper Antarc* Antarctic Wiley Online Library European Journal of Biochemistry 267 1 121 131 |
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Open Polar |
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Wiley Online Library |
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crwiley |
language |
English |
description |
The Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 contains two distinct glutamate dehydrogenases (GDH), each specific for either NADP + or NAD + . This feature is quite unusual in bacteria, which generally have a single GDH. NADP + ‐dependent GDH has been purified to homogeneity and the gene encoding GDH has been cloned and expressed. The enzyme has a hexameric structure. The amino acid sequence determined by peptide and gene analyses comprises 447 residues, yielding a protein with a molecular mass of 49 285 Da. The sequence shows homology with hexameric GDHs, with identity levels of 52% and 49% with Escherichia coli and Clostridium symbiosum GDH, respectively. The coenzyme‐binding fingerprint motif GXGXXG/A (common to all GDHs) has Ser at the last position in this enzyme. The overall hydrophilic character is increased and a five‐residue insertion in a loop between two α‐helices may contribute to the increase in protein flexibility. Psychrobacter sp. TAD1 GDH apparent temperature optimum is shifted towards low temperatures, whereas irreversible heat inactivation occurs at temperatures similar to those of E. coli GDH. The catalytic efficiency in the temperature range 10–30 °C is similar or lower than that of E. coli GDH. Unlike E. coli GDH the enzyme exhibits marked positive cooperativity towards 2‐oxoglutarate and NADPH. This feature is generally absent in prokaryotic GDHs. These observations suggest a regulatory role for this GDH, the most crucial feature being the structural/functional properties required for fine regulation of activity, rather than the high catalytic efficiency and thermolability encountered in several cold‐active enzymes. |
format |
Article in Journal/Newspaper |
author |
Di Fraia, Raffaela Wilquet, Valerie Ciardiello, M. Antonietta Carratore, Vito Antignani, Antonella Camardella, Laura Glansdorff, Nicolas di Prisco, Guido |
spellingShingle |
Di Fraia, Raffaela Wilquet, Valerie Ciardiello, M. Antonietta Carratore, Vito Antignani, Antonella Camardella, Laura Glansdorff, Nicolas di Prisco, Guido NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 |
author_facet |
Di Fraia, Raffaela Wilquet, Valerie Ciardiello, M. Antonietta Carratore, Vito Antignani, Antonella Camardella, Laura Glansdorff, Nicolas di Prisco, Guido |
author_sort |
Di Fraia, Raffaela |
title |
NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 |
title_short |
NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 |
title_full |
NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 |
title_fullStr |
NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 |
title_full_unstemmed |
NADP + ‐dependent glutamate dehydrogenase in the Antarctic psychrotolerant bacterium Psychrobacter sp. TAD1 |
title_sort |
nadp + ‐dependent glutamate dehydrogenase in the antarctic psychrotolerant bacterium psychrobacter sp. tad1 |
publisher |
Wiley |
publishDate |
2000 |
url |
http://dx.doi.org/10.1046/j.1432-1327.2000.00972.x https://api.wiley.com/onlinelibrary/tdm/v1/articles/10.1046%2Fj.1432-1327.2000.00972.x https://febs.onlinelibrary.wiley.com/doi/pdf/10.1046/j.1432-1327.2000.00972.x |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
European Journal of Biochemistry volume 267, issue 1, page 121-131 ISSN 0014-2956 1432-1033 |
op_rights |
http://onlinelibrary.wiley.com/termsAndConditions#vor |
op_doi |
https://doi.org/10.1046/j.1432-1327.2000.00972.x |
container_title |
European Journal of Biochemistry |
container_volume |
267 |
container_issue |
1 |
container_start_page |
121 |
op_container_end_page |
131 |
_version_ |
1810494357110784000 |