The co-existence of cold activity and thermal stability in an Antarctic GH42 β-galactosidase relies on its hexameric quaternary arrangement.
Amino Acid Sequence
Antarctic Regions
Bacterial Proteins
/ chemistry
Binding Sites
Cloning, Molecular
Cold Temperature
Crystallography, X-Ray
Enzyme Stability
Escherichia coli
/ genetics
Galactose
/ chemistry
Gene Expression
Genetic Vectors
/ chemistry
Kinetics
Marinomonas
/ chemistry
Models, Molecular
Phylogeny
Protein Binding
Protein Conformation, alpha-Helical
Protein Conformation, beta-Strand
Protein Interaction Domains and Motifs
Protein Multimerization
Protein Structure, Quaternary
Protein Structure, Tertiary
Recombinant Proteins
/ chemistry
Sequence Alignment
Sequence Homology, Amino Acid
Substrate Specificity
Thermodynamics
beta-Galactosidase
/ chemistry
cold adaptation
cooperativity
enzyme kinetics
glycoside hydrolase
psychrophilic enzyme
Journal
The FEBS journal
ISSN: 1742-4658
Titre abrégé: FEBS J
Pays: England
ID NLM: 101229646
Informations de publication
Date de publication:
01 2021
01 2021
Historique:
received:
03
03
2020
revised:
25
04
2020
accepted:
29
04
2020
pubmed:
5
5
2020
medline:
22
6
2021
entrez:
5
5
2020
Statut:
ppublish
Résumé
To survive in cold environments, psychrophilic organisms produce enzymes endowed with high specific activity at low temperature. The structure of these enzymes is usually flexible and mostly thermolabile. In this work, we investigate the structural basis of cold adaptation of a GH42 β-galactosidase from the psychrophilic Marinomonas ef1. This enzyme couples cold activity with astonishing robustness for a psychrophilic protein, for it retains 23% of its highest activity at 5 °C and it is stable for several days at 37 °C and even 50 °C. Phylogenetic analyses indicate a close relationship with thermophilic β-galactosidases, suggesting that the present-day enzyme evolved from a thermostable scaffold modeled by environmental selective pressure. The crystallographic structure reveals the overall similarity with GH42 enzymes, along with a hexameric arrangement (dimer of trimers) not found in psychrophilic, mesophilic, and thermophilic homologues. In the quaternary structure, protomers form a large central cavity, whose accessibility to the substrate is promoted by the dynamic behavior of surface loops, even at low temperature. A peculiar cooperative behavior of the enzyme is likely related to the increase of the internal cavity permeability triggered by heating. Overall, our results highlight a novel strategy of enzyme cold adaptation, based on the oligomerization state of the enzyme, which effectively challenges the paradigm of cold activity coupled with intrinsic thermolability. DATABASE: Structural data are available in the Protein Data Bank database under the accession number 6Y2K.
Substances chimiques
Bacterial Proteins
0
Recombinant Proteins
0
beta-Galactosidase
EC 3.2.1.23
Galactose
X2RN3Q8DNE
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
546-565Informations de copyright
© 2020 Federation of European Biochemical Societies.
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