Serial femtosecond crystallography of soluble proteins in lipidic cubic phase

Raimund Fromme, Andrii Ishchenko, Markus Metz, Shatabdi Roy Chowdhury, Shibom Basu, Sébastien Boutet, Petra Fromme, Thomas A. White, Anton Barty, John Spence, Uwe Weierstall, Wei Liu, Vadim Cherezov

    Research output: Contribution to journalArticle

    31 Citations (Scopus)

    Abstract

    Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables high-resolution protein structure determination using micrometre-sized crystals at room temperature with minimal effects from radiation damage. SFX requires a steady supply of microcrystals intersecting the XFEL beam at random orientations. An LCP-SFX method has recently been introduced in which microcrystals of membrane proteins are grown and delivered for SFX data collection inside a gel-like membrane-mimetic matrix, known as lipidic cubic phase (LCP), using a special LCP microextrusion injector. Here, it is demonstrated that LCP can also be used as a suitable carrier medium for microcrystals of soluble proteins, enabling a dramatic reduction in the amount of crystallized protein required for data collection compared with crystals delivered by liquid injectors. High-quality LCP-SFX data sets were collected for two soluble proteins, lysozyme and phycocyanin, using less than 0.1 mg of each protein.

    Original languageEnglish (US)
    Pages (from-to)545-551
    Number of pages7
    JournalIUCrJ
    Volume2
    DOIs
    StatePublished - Sep 1 2015

    Fingerprint

    Crystallography
    crystallography
    Microcrystals
    proteins
    Proteins
    microcrystals
    X ray lasers
    Free electron lasers
    injectors
    free electron lasers
    Lasers
    Phycocyanin
    X-Rays
    Electrons
    Liquid Crystals
    membranes
    Membranes
    Radiation damage
    Radiation Effects
    Muramidase

    Keywords

    • lipidic cubic phase
    • serial femtosecond crystallography
    • soluble protein
    • X-ray free-electron laser

    ASJC Scopus subject areas

    • Chemistry(all)
    • Materials Science(all)
    • Condensed Matter Physics
    • Biochemistry

    Cite this

    Fromme, R., Ishchenko, A., Metz, M., Chowdhury, S. R., Basu, S., Boutet, S., ... Cherezov, V. (2015). Serial femtosecond crystallography of soluble proteins in lipidic cubic phase. IUCrJ, 2, 545-551. https://doi.org/10.1107/S2052252515013160

    Serial femtosecond crystallography of soluble proteins in lipidic cubic phase. / Fromme, Raimund; Ishchenko, Andrii; Metz, Markus; Chowdhury, Shatabdi Roy; Basu, Shibom; Boutet, Sébastien; Fromme, Petra; White, Thomas A.; Barty, Anton; Spence, John; Weierstall, Uwe; Liu, Wei; Cherezov, Vadim.

    In: IUCrJ, Vol. 2, 01.09.2015, p. 545-551.

    Research output: Contribution to journalArticle

    Fromme, R, Ishchenko, A, Metz, M, Chowdhury, SR, Basu, S, Boutet, S, Fromme, P, White, TA, Barty, A, Spence, J, Weierstall, U, Liu, W & Cherezov, V 2015, 'Serial femtosecond crystallography of soluble proteins in lipidic cubic phase' IUCrJ, vol. 2, pp. 545-551. https://doi.org/10.1107/S2052252515013160
    Fromme, Raimund ; Ishchenko, Andrii ; Metz, Markus ; Chowdhury, Shatabdi Roy ; Basu, Shibom ; Boutet, Sébastien ; Fromme, Petra ; White, Thomas A. ; Barty, Anton ; Spence, John ; Weierstall, Uwe ; Liu, Wei ; Cherezov, Vadim. / Serial femtosecond crystallography of soluble proteins in lipidic cubic phase. In: IUCrJ. 2015 ; Vol. 2. pp. 545-551.
    @article{49043a8a0e754b1d81855b908b6dca9c,
    title = "Serial femtosecond crystallography of soluble proteins in lipidic cubic phase",
    abstract = "Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables high-resolution protein structure determination using micrometre-sized crystals at room temperature with minimal effects from radiation damage. SFX requires a steady supply of microcrystals intersecting the XFEL beam at random orientations. An LCP-SFX method has recently been introduced in which microcrystals of membrane proteins are grown and delivered for SFX data collection inside a gel-like membrane-mimetic matrix, known as lipidic cubic phase (LCP), using a special LCP microextrusion injector. Here, it is demonstrated that LCP can also be used as a suitable carrier medium for microcrystals of soluble proteins, enabling a dramatic reduction in the amount of crystallized protein required for data collection compared with crystals delivered by liquid injectors. High-quality LCP-SFX data sets were collected for two soluble proteins, lysozyme and phycocyanin, using less than 0.1 mg of each protein.",
    keywords = "lipidic cubic phase, serial femtosecond crystallography, soluble protein, X-ray free-electron laser",
    author = "Raimund Fromme and Andrii Ishchenko and Markus Metz and Chowdhury, {Shatabdi Roy} and Shibom Basu and S{\'e}bastien Boutet and Petra Fromme and White, {Thomas A.} and Anton Barty and John Spence and Uwe Weierstall and Wei Liu and Vadim Cherezov",
    year = "2015",
    month = "9",
    day = "1",
    doi = "10.1107/S2052252515013160",
    language = "English (US)",
    volume = "2",
    pages = "545--551",
    journal = "IUCrJ",
    issn = "2052-2525",
    publisher = "International Union of Crystallography",

    }

    TY - JOUR

    T1 - Serial femtosecond crystallography of soluble proteins in lipidic cubic phase

    AU - Fromme, Raimund

    AU - Ishchenko, Andrii

    AU - Metz, Markus

    AU - Chowdhury, Shatabdi Roy

    AU - Basu, Shibom

    AU - Boutet, Sébastien

    AU - Fromme, Petra

    AU - White, Thomas A.

    AU - Barty, Anton

    AU - Spence, John

    AU - Weierstall, Uwe

    AU - Liu, Wei

    AU - Cherezov, Vadim

    PY - 2015/9/1

    Y1 - 2015/9/1

    N2 - Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables high-resolution protein structure determination using micrometre-sized crystals at room temperature with minimal effects from radiation damage. SFX requires a steady supply of microcrystals intersecting the XFEL beam at random orientations. An LCP-SFX method has recently been introduced in which microcrystals of membrane proteins are grown and delivered for SFX data collection inside a gel-like membrane-mimetic matrix, known as lipidic cubic phase (LCP), using a special LCP microextrusion injector. Here, it is demonstrated that LCP can also be used as a suitable carrier medium for microcrystals of soluble proteins, enabling a dramatic reduction in the amount of crystallized protein required for data collection compared with crystals delivered by liquid injectors. High-quality LCP-SFX data sets were collected for two soluble proteins, lysozyme and phycocyanin, using less than 0.1 mg of each protein.

    AB - Serial femtosecond crystallography (SFX) at X-ray free-electron lasers (XFELs) enables high-resolution protein structure determination using micrometre-sized crystals at room temperature with minimal effects from radiation damage. SFX requires a steady supply of microcrystals intersecting the XFEL beam at random orientations. An LCP-SFX method has recently been introduced in which microcrystals of membrane proteins are grown and delivered for SFX data collection inside a gel-like membrane-mimetic matrix, known as lipidic cubic phase (LCP), using a special LCP microextrusion injector. Here, it is demonstrated that LCP can also be used as a suitable carrier medium for microcrystals of soluble proteins, enabling a dramatic reduction in the amount of crystallized protein required for data collection compared with crystals delivered by liquid injectors. High-quality LCP-SFX data sets were collected for two soluble proteins, lysozyme and phycocyanin, using less than 0.1 mg of each protein.

    KW - lipidic cubic phase

    KW - serial femtosecond crystallography

    KW - soluble protein

    KW - X-ray free-electron laser

    UR - http://www.scopus.com/inward/record.url?scp=84940932911&partnerID=8YFLogxK

    UR - http://www.scopus.com/inward/citedby.url?scp=84940932911&partnerID=8YFLogxK

    U2 - 10.1107/S2052252515013160

    DO - 10.1107/S2052252515013160

    M3 - Article

    VL - 2

    SP - 545

    EP - 551

    JO - IUCrJ

    JF - IUCrJ

    SN - 2052-2525

    ER -