Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18
peer reviewed The gene encoding the phosphoglycerate kinase (PGK) from the Antarctic Pseudomonas sp. TACII18 has been cloned and found to be inserted between the genes encoding for glyceraldhyde-3-phosphate dehydrogenase and fructose aldolase. The His-tagged and the native recombinant PGK from the p...
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American Society for Biochemistry and Molecular Biology
2000
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ftorbi:oai:orbi.ulg.ac.be:2268/16106 2024-04-21T07:49:42+00:00 Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 Bentahir, Mostafa Feller, Georges Aittaleb, Mohamed Lamotte-Brasseur, Josette Himri, Touhami Chessa, Jean-Pierre Gerday, Charles 2000-04-14 https://orbi.uliege.be/handle/2268/16106 https://orbi.uliege.be/bitstream/2268/16106/1/JBC_2000_PGK.pdf https://doi.org/10.1074/jbc.275.15.11147 en eng American Society for Biochemistry and Molecular Biology urn:issn:0021-9258 urn:issn:1083-351X https://orbi.uliege.be/handle/2268/16106 info:hdl:2268/16106 https://orbi.uliege.be/bitstream/2268/16106/1/JBC_2000_PGK.pdf doi:10.1074/jbc.275.15.11147 scopus-id:2-s2.0-0034646605 info:pmid:10753921 open access http://purl.org/coar/access_right/c_abf2 info:eu-repo/semantics/openAccess Journal of Biological Chemistry, 275 (15), 11147-53 (2000-04-14) 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 2000 ftorbi https://doi.org/10.1074/jbc.275.15.11147 2024-03-27T14:55:11Z peer reviewed The gene encoding the phosphoglycerate kinase (PGK) from the Antarctic Pseudomonas sp. TACII18 has been cloned and found to be inserted between the genes encoding for glyceraldhyde-3-phosphate dehydrogenase and fructose aldolase. The His-tagged and the native recombinant PGK from the psychrophilic Pseudomonas were expressed in Escherichia coli. The wild-type and the native recombinant enzymes displayed identical properties, such as a decreased thermostability and a 2-fold higher catalytic efficiency at 25 degrees C when compared with the mesophilic PGK from yeast. These properties, which reflect typical features of cold-adapted enzymes, were strongly altered in the His-tagged recombinant PGK. The structural model of the psychrophilic PGK indicated that a key determinant of its low stability is the reduced number of salt bridges, surface charges, and aromatic interactions when compared with mesophilic and thermophilic PGK. Differential scanning calorimetry of the psychrophilic PGK revealed unusual variations in its conformational stability for the free and substrate-bound forms. In the free form, a heat-labile and a thermostable domain unfold independently. It is proposed that the heat-labile domain acts as a destabilizing domain, providing the required flexibility around the active site for catalysis at low temperatures. Article in Journal/Newspaper Antarc* Antarctic University of Liège: ORBi (Open Repository and Bibliography) Journal of Biological Chemistry 275 15 11147 11153 |
institution |
Open Polar |
collection |
University of Liège: ORBi (Open Repository and Bibliography) |
op_collection_id |
ftorbi |
language |
English |
topic |
Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
spellingShingle |
Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire Bentahir, Mostafa Feller, Georges Aittaleb, Mohamed Lamotte-Brasseur, Josette Himri, Touhami Chessa, Jean-Pierre Gerday, Charles Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 |
topic_facet |
Life sciences Biochemistry biophysics & molecular biology Sciences du vivant Biochimie biophysique & biologie moléculaire |
description |
peer reviewed The gene encoding the phosphoglycerate kinase (PGK) from the Antarctic Pseudomonas sp. TACII18 has been cloned and found to be inserted between the genes encoding for glyceraldhyde-3-phosphate dehydrogenase and fructose aldolase. The His-tagged and the native recombinant PGK from the psychrophilic Pseudomonas were expressed in Escherichia coli. The wild-type and the native recombinant enzymes displayed identical properties, such as a decreased thermostability and a 2-fold higher catalytic efficiency at 25 degrees C when compared with the mesophilic PGK from yeast. These properties, which reflect typical features of cold-adapted enzymes, were strongly altered in the His-tagged recombinant PGK. The structural model of the psychrophilic PGK indicated that a key determinant of its low stability is the reduced number of salt bridges, surface charges, and aromatic interactions when compared with mesophilic and thermophilic PGK. Differential scanning calorimetry of the psychrophilic PGK revealed unusual variations in its conformational stability for the free and substrate-bound forms. In the free form, a heat-labile and a thermostable domain unfold independently. It is proposed that the heat-labile domain acts as a destabilizing domain, providing the required flexibility around the active site for catalysis at low temperatures. |
format |
Article in Journal/Newspaper |
author |
Bentahir, Mostafa Feller, Georges Aittaleb, Mohamed Lamotte-Brasseur, Josette Himri, Touhami Chessa, Jean-Pierre Gerday, Charles |
author_facet |
Bentahir, Mostafa Feller, Georges Aittaleb, Mohamed Lamotte-Brasseur, Josette Himri, Touhami Chessa, Jean-Pierre Gerday, Charles |
author_sort |
Bentahir, Mostafa |
title |
Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 |
title_short |
Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 |
title_full |
Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 |
title_fullStr |
Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 |
title_full_unstemmed |
Structural, Kinetic, and Calorimetric Characterization of the Cold-Active Phosphoglycerate Kinase from the Antarctic Pseudomonas Sp. Tacii18 |
title_sort |
structural, kinetic, and calorimetric characterization of the cold-active phosphoglycerate kinase from the antarctic pseudomonas sp. tacii18 |
publisher |
American Society for Biochemistry and Molecular Biology |
publishDate |
2000 |
url |
https://orbi.uliege.be/handle/2268/16106 https://orbi.uliege.be/bitstream/2268/16106/1/JBC_2000_PGK.pdf https://doi.org/10.1074/jbc.275.15.11147 |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_source |
Journal of Biological Chemistry, 275 (15), 11147-53 (2000-04-14) |
op_relation |
urn:issn:0021-9258 urn:issn:1083-351X https://orbi.uliege.be/handle/2268/16106 info:hdl:2268/16106 https://orbi.uliege.be/bitstream/2268/16106/1/JBC_2000_PGK.pdf doi:10.1074/jbc.275.15.11147 scopus-id:2-s2.0-0034646605 info:pmid:10753921 |
op_rights |
open access http://purl.org/coar/access_right/c_abf2 info:eu-repo/semantics/openAccess |
op_doi |
https://doi.org/10.1074/jbc.275.15.11147 |
container_title |
Journal of Biological Chemistry |
container_volume |
275 |
container_issue |
15 |
container_start_page |
11147 |
op_container_end_page |
11153 |
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1796933717094039552 |