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

33 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

AN - SCOPUS:84940932911

VL - 2

SP - 545

EP - 551

JO - IUCrJ

JF - IUCrJ

SN - 2052-2525

ER -