The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium.
International audience The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by...
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Online Access: | https://hal.science/hal-00473856 https://doi.org/10.1042/BJ20070640 |
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ftinstpasteur:oai:HAL:hal-00473856v1 2024-05-19T07:31:33+00:00 The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles Interactions et Modulateurs de Réponses (IMR) Centre National de la Recherche Scientifique (CNRS) Centre d’Ingénierie des Protéines Université de Liège = Centre for Protein Engineering University of Liège (CIP) Université de Liège Résonance Magnétique Nucléaire des Biomolécules Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS) Institut de biologie et chimie des protéines Lyon (IBCP) Université Claude Bernard Lyon 1 (UCBL) Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS) 2007-10-15 https://hal.science/hal-00473856 https://doi.org/10.1042/BJ20070640 en eng HAL CCSD Portland Press info:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20070640 info:eu-repo/semantics/altIdentifier/pmid/17635108 hal-00473856 https://hal.science/hal-00473856 doi:10.1042/BJ20070640 PUBMED: 17635108 PUBMEDCENTRAL: PMC2049020 ISSN: 0264-6021 EISSN: 1470-8728 Biochemical Journal https://hal.science/hal-00473856 Biochemical Journal, 2007, 407 (2), pp.293-302. ⟨10.1042/BJ20070640⟩ MESH: Acclimatization MESH: Catalysis MESH: Cellulase MESH: Cold Temperature MESH: Enzyme Stability MESH: Mutation MESH: Protein Conformation MESH: Pseudoalteromonas [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology info:eu-repo/semantics/article Journal articles 2007 ftinstpasteur https://doi.org/10.1042/BJ20070640 2024-04-24T23:56:18Z International audience The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by three disulfide bridges. To evaluate the possible role of this region in cold adaptation, the LR was sequentially shortened by protein engineering, successively deleting one and two loops of this module, whereas the last disulfide bridge was also suppressed by replacing the last two cysteine residue by two alanine residues. The kinetic and thermodynamic properties of the mutants were compared with those of the full-length enzyme, and also with those of the cold-adapted CM alone and with those of the homologous mesophilic enzyme, Cel5A, from Erwinia chrysanthemi. The thermostability of the mutated enzymes as well as their relative flexibility were evaluated by differential scanning calorimetry and fluorescence quenching respectively. The topology of the structure of the shortest mutant was determined by SAXS (small-angle X-ray scattering). The data indicate that the sequential shortening of the LR induces a regular decrease of the specific activity towards macromolecular substrates, reduces the relative flexibility and concomitantly increases the thermostability of the shortened enzymes. This demonstrates that the long LR of the full-length enzyme favours the catalytic efficiency at low and moderate temperatures by rendering the structure not only less compact, but also less stable, and plays a crucial role in the adaptation to cold of this cellulolytic enzyme. Article in Journal/Newspaper Antarc* Antarctic Institut Pasteur: HAL Biochemical Journal 407 2 293 302 |
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Open Polar |
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Institut Pasteur: HAL |
op_collection_id |
ftinstpasteur |
language |
English |
topic |
MESH: Acclimatization MESH: Catalysis MESH: Cellulase MESH: Cold Temperature MESH: Enzyme Stability MESH: Mutation MESH: Protein Conformation MESH: Pseudoalteromonas [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology |
spellingShingle |
MESH: Acclimatization MESH: Catalysis MESH: Cellulase MESH: Cold Temperature MESH: Enzyme Stability MESH: Mutation MESH: Protein Conformation MESH: Pseudoalteromonas [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. |
topic_facet |
MESH: Acclimatization MESH: Catalysis MESH: Cellulase MESH: Cold Temperature MESH: Enzyme Stability MESH: Mutation MESH: Protein Conformation MESH: Pseudoalteromonas [SDV.BBM]Life Sciences [q-bio]/Biochemistry Molecular Biology |
description |
International audience The psychrophilic cellulase, Cel5G, from the Antarctic bacterium Pseudoalteromonas haloplanktis is composed of a catalytic module (CM) joined to a carbohydrate-binding module (CBM) by an unusually long, extended and flexible linker region (LR) containing three loops closed by three disulfide bridges. To evaluate the possible role of this region in cold adaptation, the LR was sequentially shortened by protein engineering, successively deleting one and two loops of this module, whereas the last disulfide bridge was also suppressed by replacing the last two cysteine residue by two alanine residues. The kinetic and thermodynamic properties of the mutants were compared with those of the full-length enzyme, and also with those of the cold-adapted CM alone and with those of the homologous mesophilic enzyme, Cel5A, from Erwinia chrysanthemi. The thermostability of the mutated enzymes as well as their relative flexibility were evaluated by differential scanning calorimetry and fluorescence quenching respectively. The topology of the structure of the shortest mutant was determined by SAXS (small-angle X-ray scattering). The data indicate that the sequential shortening of the LR induces a regular decrease of the specific activity towards macromolecular substrates, reduces the relative flexibility and concomitantly increases the thermostability of the shortened enzymes. This demonstrates that the long LR of the full-length enzyme favours the catalytic efficiency at low and moderate temperatures by rendering the structure not only less compact, but also less stable, and plays a crucial role in the adaptation to cold of this cellulolytic enzyme. |
author2 |
Interactions et Modulateurs de Réponses (IMR) Centre National de la Recherche Scientifique (CNRS) Centre d’Ingénierie des Protéines Université de Liège = Centre for Protein Engineering University of Liège (CIP) Université de Liège Résonance Magnétique Nucléaire des Biomolécules Institut Pasteur Paris (IP)-Centre National de la Recherche Scientifique (CNRS) Institut de biologie et chimie des protéines Lyon (IBCP) Université Claude Bernard Lyon 1 (UCBL) Université de Lyon-Université de Lyon-Centre National de la Recherche Scientifique (CNRS) |
format |
Article in Journal/Newspaper |
author |
Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles |
author_facet |
Sonan, Guillaume K Receveur-Brechot, Véronique Duez, Colette Aghajari, Nushin Czjzek, Mirjam Haser, Richard Gerday, Charles |
author_sort |
Sonan, Guillaume K |
title |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. |
title_short |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. |
title_full |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. |
title_fullStr |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. |
title_full_unstemmed |
The linker region plays a key role in the adaptation to cold of the cellulase from an Antarctic bacterium. |
title_sort |
linker region plays a key role in the adaptation to cold of the cellulase from an antarctic bacterium. |
publisher |
HAL CCSD |
publishDate |
2007 |
url |
https://hal.science/hal-00473856 https://doi.org/10.1042/BJ20070640 |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
ISSN: 0264-6021 EISSN: 1470-8728 Biochemical Journal https://hal.science/hal-00473856 Biochemical Journal, 2007, 407 (2), pp.293-302. ⟨10.1042/BJ20070640⟩ |
op_relation |
info:eu-repo/semantics/altIdentifier/doi/10.1042/BJ20070640 info:eu-repo/semantics/altIdentifier/pmid/17635108 hal-00473856 https://hal.science/hal-00473856 doi:10.1042/BJ20070640 PUBMED: 17635108 PUBMEDCENTRAL: PMC2049020 |
op_doi |
https://doi.org/10.1042/BJ20070640 |
container_title |
Biochemical Journal |
container_volume |
407 |
container_issue |
2 |
container_start_page |
293 |
op_container_end_page |
302 |
_version_ |
1799469411830071296 |