Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering
peer reviewed Pseudoalteromonas haloplanktis is a psychrophilic Gram-negative bacterium isolated in Antarctica, that lives on organic remains of algae. This bacterium converts the cellulose, highly constitutive of algae, into an immediate nutritive form by biodegrading this biopolymer. To understand...
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2005
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Online Access: | https://orbi.uliege.be/handle/2268/15357 https://doi.org/10.1016/j.jmb.2005.03.026 |
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ftorbi:oai:orbi.ulg.ac.be:2268/15357 2024-04-21T07:51:17+00:00 Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering Violot, S. Aghajari, N. Czjzek, M. Feller, Georges Sonan, G. K. Gouet, P. Gerday, Charles Haser, R. Receveur-Brechot, V. 2005-05-20 https://orbi.uliege.be/handle/2268/15357 https://doi.org/10.1016/j.jmb.2005.03.026 en eng Academic Press Ltd Elsevier Science Ltd urn:issn:0022-2836 urn:issn:1089-8638 https://orbi.uliege.be/handle/2268/15357 info:hdl:2268/15357 doi:10.1016/j.jmb.2005.03.026 scopus-id:2-s2.0-18044372844 info:pmid:15854656 restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess Journal of Molecular Biology, 348 (5), 1211-1224 (2005-05-20) cold adaptation protein disorder cellulose protein flexibility TSP3 Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire journal article http://purl.org/coar/resource_type/c_6501 info:eu-repo/semantics/article peer reviewed 2005 ftorbi https://doi.org/10.1016/j.jmb.2005.03.026 2024-03-27T14:54:55Z peer reviewed Pseudoalteromonas haloplanktis is a psychrophilic Gram-negative bacterium isolated in Antarctica, that lives on organic remains of algae. This bacterium converts the cellulose, highly constitutive of algae, into an immediate nutritive form by biodegrading this biopolymer. To understand the mechanisms of cold adaptation of its enzymatic components, we studied the structural properties of an endoglucanase, Cel5G, by complementary methods, X-ray crystallography and small angle X-ray scattering. Using X-ray crystallography, we determined the structure of the catalytic core module of this family 5 endoglucanase, at 1.4 angstrom resolution in its native form and at 1.6 angstrom in the cellobiose-bound form. The catalytic module of Cel5G presents the (beta/alpha)(8)-barrel structure typical of clan GH-A of glycoside hydrolase families. The structural comparison of the catalytic core of Cel5G with the mesophilic catalytic core of Cel5A from Erwinia chrysanthemi revealed modifications at the atomic level leading to higher flexibility and thermolability, which might account for the higher activity of Cel5G at low temperatures. Using small angle X-ray scattering we further explored the structure at the entire enzyme level. We analyzed the dimensions, shape, and conformation of Cel5G full length in solution and especially of the linker between the catalytic module and the cellulose-binding module. The results showed that the linker is unstructured, and unusually long and flexible, a peculiarity that distinguishes it from its mesophilic counterpart. Loops formed at the base by disulfide bridges presumably add constraints to stabilize the most extended conformations. These results suggest that the linker plays a major role in cold adaptation of this psychrophilic enzyme, allowing steric optimization of substrate accessibility. (c) 2005 Elsevier Ltd. All rights reserved. Article in Journal/Newspaper Antarc* Antarctica University of Liège: ORBi (Open Repository and Bibliography) Journal of Molecular Biology 348 5 1211 1224 |
institution |
Open Polar |
collection |
University of Liège: ORBi (Open Repository and Bibliography) |
op_collection_id |
ftorbi |
language |
English |
topic |
cold adaptation protein disorder cellulose protein flexibility TSP3 Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
spellingShingle |
cold adaptation protein disorder cellulose protein flexibility TSP3 Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire Violot, S. Aghajari, N. Czjzek, M. Feller, Georges Sonan, G. K. Gouet, P. Gerday, Charles Haser, R. Receveur-Brechot, V. Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering |
topic_facet |
cold adaptation protein disorder cellulose protein flexibility TSP3 Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
description |
peer reviewed Pseudoalteromonas haloplanktis is a psychrophilic Gram-negative bacterium isolated in Antarctica, that lives on organic remains of algae. This bacterium converts the cellulose, highly constitutive of algae, into an immediate nutritive form by biodegrading this biopolymer. To understand the mechanisms of cold adaptation of its enzymatic components, we studied the structural properties of an endoglucanase, Cel5G, by complementary methods, X-ray crystallography and small angle X-ray scattering. Using X-ray crystallography, we determined the structure of the catalytic core module of this family 5 endoglucanase, at 1.4 angstrom resolution in its native form and at 1.6 angstrom in the cellobiose-bound form. The catalytic module of Cel5G presents the (beta/alpha)(8)-barrel structure typical of clan GH-A of glycoside hydrolase families. The structural comparison of the catalytic core of Cel5G with the mesophilic catalytic core of Cel5A from Erwinia chrysanthemi revealed modifications at the atomic level leading to higher flexibility and thermolability, which might account for the higher activity of Cel5G at low temperatures. Using small angle X-ray scattering we further explored the structure at the entire enzyme level. We analyzed the dimensions, shape, and conformation of Cel5G full length in solution and especially of the linker between the catalytic module and the cellulose-binding module. The results showed that the linker is unstructured, and unusually long and flexible, a peculiarity that distinguishes it from its mesophilic counterpart. Loops formed at the base by disulfide bridges presumably add constraints to stabilize the most extended conformations. These results suggest that the linker plays a major role in cold adaptation of this psychrophilic enzyme, allowing steric optimization of substrate accessibility. (c) 2005 Elsevier Ltd. All rights reserved. |
format |
Article in Journal/Newspaper |
author |
Violot, S. Aghajari, N. Czjzek, M. Feller, Georges Sonan, G. K. Gouet, P. Gerday, Charles Haser, R. Receveur-Brechot, V. |
author_facet |
Violot, S. Aghajari, N. Czjzek, M. Feller, Georges Sonan, G. K. Gouet, P. Gerday, Charles Haser, R. Receveur-Brechot, V. |
author_sort |
Violot, S. |
title |
Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering |
title_short |
Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering |
title_full |
Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering |
title_fullStr |
Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering |
title_full_unstemmed |
Structure of a full length psychrophilic cellulase from Pseudoalteromonas haloplanktis revealed by X-ray diffraction and small angle X-ray scattering |
title_sort |
structure of a full length psychrophilic cellulase from pseudoalteromonas haloplanktis revealed by x-ray diffraction and small angle x-ray scattering |
publisher |
Academic Press Ltd Elsevier Science Ltd |
publishDate |
2005 |
url |
https://orbi.uliege.be/handle/2268/15357 https://doi.org/10.1016/j.jmb.2005.03.026 |
genre |
Antarc* Antarctica |
genre_facet |
Antarc* Antarctica |
op_source |
Journal of Molecular Biology, 348 (5), 1211-1224 (2005-05-20) |
op_relation |
urn:issn:0022-2836 urn:issn:1089-8638 https://orbi.uliege.be/handle/2268/15357 info:hdl:2268/15357 doi:10.1016/j.jmb.2005.03.026 scopus-id:2-s2.0-18044372844 info:pmid:15854656 |
op_rights |
restricted access http://purl.org/coar/access_right/c_16ec info:eu-repo/semantics/restrictedAccess |
op_doi |
https://doi.org/10.1016/j.jmb.2005.03.026 |
container_title |
Journal of Molecular Biology |
container_volume |
348 |
container_issue |
5 |
container_start_page |
1211 |
op_container_end_page |
1224 |
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1796934694853410816 |