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Atomic resolution of the crystal structure of the hyperthermophilic family 12 endocellulase and stabilizing role of the DxDxDG calcium-binding motif in Pyrococcus furiosus.

Abstract

Hyperthermophilic glycoside hydrolase family 12 endocellulase (EGPf) from the archaeon Pyrococcus furiosus catalyzes the hydrolytic cleavage of β-1,4-glucosidic linkage in β-glucan cellulose. A truncated EGPf (EGPfΔN30) mutant lacking the proline and hydroxyl-residue rich region at the N terminus was constructed, and its crystal structure was resolved at an atomic resolution of 1.07 Å. Our results indicate that the structure of EGPf, which consists of a β-jelly roll, exhibits structural similarity with the endocellulase of Thermotoga maritima. Additionally, we further determined that the thermostability of EGPf is maintained in part by the binding of Ca²⁺ in a DxDxDG Ca²⁺-binding motif, atypical of most archaeal proteins.

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  • Publisher Full Text

    • Authors

    Kim HW, Kataoka M, Ishikawa K

    Institution

    National Institute of Advanced Industrial Science and Technology, Biomass Technology Research Center, 3-11-32 Kagamiyama, Higashi-Hiroshima, Hiroshima 739-0046, Japan.

    Source

    FEBS letters 586:7 2012 Apr 5 pg 1009-13

    MeSH

    Amino Acid Motifs
    Archaeal Proteins
    Binding Sites
    Calcium
    Calorimetry, Differential Scanning
    Catalytic Domain
    Cellulases
    Cellulose
    Codon, Nonsense
    Crystallography, X-Ray
    Enzyme Stability
    Hot Temperature
    Isoenzymes
    Ligands
    Models, Molecular
    Peptide Fragments
    Protein Conformation
    Pyrococcus furiosus
    Recombinant Proteins
    beta-Glucans

    Pub Type(s)

    Journal Article

    Language

    eng

    PubMed ID

    22569255