Purification, crystallization and preliminary X-ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus

Mayumi Igura, Nobuo Maita, Takayuki Obita, Jun Kamishikiryo, Katsumi Maenaka, Daisuke Kohda

    Research output: Contribution to journalArticle

    12 Citations (Scopus)

    Abstract

    Oligosaccharyltransferase catalyzes the transfer of preassembled oligosaccharides onto asparagine residues in nascent polypeptide chains. The STT3 subunit is thought to bear the catalytic site. The C-terminal domain of the STT3 protein of Pyrococcus furiosus was expressed in Escherichia coli cells. STT3 protein prepared from two different sources, the soluble fraction and the inclusion bodies, produced crystals that diffracted to 2.7 Å. During crystallization screening, cocrystals of P. furiosus STT3 with an E. coli 50S ribosomal protein, L7/L12, were accidentally obtained. This cross-species interaction is not biologically relevant, but may be used to design a built-in polypeptide substrate for the STT3 crystals.

    Original languageEnglish
    Pages (from-to)798-801
    Number of pages4
    JournalActa Crystallographica Section F: Structural Biology and Crystallization Communications
    Volume63
    Issue number9
    DOIs
    Publication statusPublished - Aug 10 2007

    Fingerprint

    Pyrococcus furiosus
    Archaea
    Crystallization
    purification
    X-Ray Diffraction
    Escherichia coli
    Purification
    polypeptides
    crystallization
    proteins
    X ray diffraction
    Peptides
    Crystals
    Asparagine
    Inclusion Bodies
    Oligosaccharides
    diffraction
    Catalytic Domain
    Screening
    Proteins

    All Science Journal Classification (ASJC) codes

    • Biophysics
    • Structural Biology
    • Biochemistry
    • Genetics
    • Condensed Matter Physics

    Cite this

    @article{0c1311b983b849c88aaadba995094a55,
    title = "Purification, crystallization and preliminary X-ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus",
    abstract = "Oligosaccharyltransferase catalyzes the transfer of preassembled oligosaccharides onto asparagine residues in nascent polypeptide chains. The STT3 subunit is thought to bear the catalytic site. The C-terminal domain of the STT3 protein of Pyrococcus furiosus was expressed in Escherichia coli cells. STT3 protein prepared from two different sources, the soluble fraction and the inclusion bodies, produced crystals that diffracted to 2.7 {\AA}. During crystallization screening, cocrystals of P. furiosus STT3 with an E. coli 50S ribosomal protein, L7/L12, were accidentally obtained. This cross-species interaction is not biologically relevant, but may be used to design a built-in polypeptide substrate for the STT3 crystals.",
    author = "Mayumi Igura and Nobuo Maita and Takayuki Obita and Jun Kamishikiryo and Katsumi Maenaka and Daisuke Kohda",
    year = "2007",
    month = "8",
    day = "10",
    doi = "10.1107/S1744309107040134",
    language = "English",
    volume = "63",
    pages = "798--801",
    journal = "Acta Crystallographica Section F:Structural Biology Communications",
    issn = "1744-3091",
    publisher = "John Wiley and Sons Ltd",
    number = "9",

    }

    TY - JOUR

    T1 - Purification, crystallization and preliminary X-ray diffraction studies of the soluble domain of the oligosaccharyltransferase STT3 subunit from the thermophilic archaeon Pyrococcus furiosus

    AU - Igura, Mayumi

    AU - Maita, Nobuo

    AU - Obita, Takayuki

    AU - Kamishikiryo, Jun

    AU - Maenaka, Katsumi

    AU - Kohda, Daisuke

    PY - 2007/8/10

    Y1 - 2007/8/10

    N2 - Oligosaccharyltransferase catalyzes the transfer of preassembled oligosaccharides onto asparagine residues in nascent polypeptide chains. The STT3 subunit is thought to bear the catalytic site. The C-terminal domain of the STT3 protein of Pyrococcus furiosus was expressed in Escherichia coli cells. STT3 protein prepared from two different sources, the soluble fraction and the inclusion bodies, produced crystals that diffracted to 2.7 Å. During crystallization screening, cocrystals of P. furiosus STT3 with an E. coli 50S ribosomal protein, L7/L12, were accidentally obtained. This cross-species interaction is not biologically relevant, but may be used to design a built-in polypeptide substrate for the STT3 crystals.

    AB - Oligosaccharyltransferase catalyzes the transfer of preassembled oligosaccharides onto asparagine residues in nascent polypeptide chains. The STT3 subunit is thought to bear the catalytic site. The C-terminal domain of the STT3 protein of Pyrococcus furiosus was expressed in Escherichia coli cells. STT3 protein prepared from two different sources, the soluble fraction and the inclusion bodies, produced crystals that diffracted to 2.7 Å. During crystallization screening, cocrystals of P. furiosus STT3 with an E. coli 50S ribosomal protein, L7/L12, were accidentally obtained. This cross-species interaction is not biologically relevant, but may be used to design a built-in polypeptide substrate for the STT3 crystals.

    UR - http://www.scopus.com/inward/record.url?scp=34548405722&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=34548405722&partnerID=8YFLogxK

    U2 - 10.1107/S1744309107040134

    DO - 10.1107/S1744309107040134

    M3 - Article

    C2 - 17768359

    AN - SCOPUS:34548405722

    VL - 63

    SP - 798

    EP - 801

    JO - Acta Crystallographica Section F:Structural Biology Communications

    JF - Acta Crystallographica Section F:Structural Biology Communications

    SN - 1744-3091

    IS - 9

    ER -