Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin

Kehan Xu; Emil Dedic; Patricia Cob-Cantal; Christian Dienemann; Andreas Boggild; Kristoffer S. Winther; Kenn Gerdes; Ditlev E. Brodersen

doi:10.1107/S1744309113014012

Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin

Kehan Xu, Emil Dedic, Patricia Cob-Cantal, Christian Dienemann, Andreas Boggild, Kristoffer S. Winther, Kenn Gerdes, Ditlev E. Brodersen

6 Citations (Scopus)

Abstract

Upon release from the stable complex formed with its antitoxin VapB, the toxin VapC (MvpT) of the Gram-negative pathogen Shigella flexneri is capable of globally down-regulating translation by specifically cleaving initiator tRNA^fMet in the anticodon region. Recombinant Shigella flexneri VapC^D7A harbouring an active-site mutation was overexpressed in Escherichia coli, purified to homogeneity and crystallized by the vapour-diffusion technique. A preliminary X-ray crystallographic analysis shows that the crystals diffracted to at least 1.9 Å resolution at a synchrotron X-ray source and belonged to the trigonal space group in the hexagonal setting, H3, with unit-cell parameters a = b = 120.1, c = 52.5 Å, α = β = 90, γ = 120°. The Matthews coefficient is 2.46 Å³ Da^-1, suggesting two molecules per asymmetric unit and corresponding to a solvent content of 50.0%.

Original language	English
Journal	Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online
Volume	69
Issue number	7
Pages (from-to)	762-765
Number of pages	4
ISSN	1744-3091
DOIs	https://doi.org/10.1107/S1744309113014012
Publication status	Published - Jul 2013
Externally published	Yes

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Xu, K., Dedic, E., Cob-Cantal, P., Dienemann, C., Boggild, A., Winther, K. S., Gerdes, K., & Brodersen, D. E. (2013). Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin. Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online, 69(7), 762-765. https://doi.org/10.1107/S1744309113014012

Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin. / Xu, Kehan; Dedic, Emil; Cob-Cantal, Patricia et al.

In: Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online, Vol. 69, No. 7, 07.2013, p. 762-765.

Research output: Contribution to journal › Journal article › Research › peer-review

Xu, K, Dedic, E, Cob-Cantal, P, Dienemann, C, Boggild, A, Winther, KS, Gerdes, K & Brodersen, DE 2013, 'Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin', Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online, vol. 69, no. 7, pp. 762-765. https://doi.org/10.1107/S1744309113014012

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title = "Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin",

abstract = "Upon release from the stable complex formed with its antitoxin VapB, the toxin VapC (MvpT) of the Gram-negative pathogen Shigella flexneri is capable of globally down-regulating translation by specifically cleaving initiator tRNAfMet in the anticodon region. Recombinant Shigella flexneri VapCD7A harbouring an active-site mutation was overexpressed in Escherichia coli, purified to homogeneity and crystallized by the vapour-diffusion technique. A preliminary X-ray crystallographic analysis shows that the crystals diffracted to at least 1.9 {\AA} resolution at a synchrotron X-ray source and belonged to the trigonal space group in the hexagonal setting, H3, with unit-cell parameters a = b = 120.1, c = 52.5 {\AA}, α = β = 90, γ = 120°. The Matthews coefficient is 2.46 {\AA}3 Da-1, suggesting two molecules per asymmetric unit and corresponding to a solvent content of 50.0%.",

author = "Kehan Xu and Emil Dedic and Patricia Cob-Cantal and Christian Dienemann and Andreas Boggild and Winther, {Kristoffer S.} and Kenn Gerdes and Brodersen, {Ditlev E.}",

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T1 - Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin

AU - Xu, Kehan

AU - Dedic, Emil

AU - Cob-Cantal, Patricia

AU - Dienemann, Christian

AU - Boggild, Andreas

AU - Winther, Kristoffer S.

AU - Gerdes, Kenn

AU - Brodersen, Ditlev E.

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N2 - Upon release from the stable complex formed with its antitoxin VapB, the toxin VapC (MvpT) of the Gram-negative pathogen Shigella flexneri is capable of globally down-regulating translation by specifically cleaving initiator tRNAfMet in the anticodon region. Recombinant Shigella flexneri VapCD7A harbouring an active-site mutation was overexpressed in Escherichia coli, purified to homogeneity and crystallized by the vapour-diffusion technique. A preliminary X-ray crystallographic analysis shows that the crystals diffracted to at least 1.9 Å resolution at a synchrotron X-ray source and belonged to the trigonal space group in the hexagonal setting, H3, with unit-cell parameters a = b = 120.1, c = 52.5 Å, α = β = 90, γ = 120°. The Matthews coefficient is 2.46 Å3 Da-1, suggesting two molecules per asymmetric unit and corresponding to a solvent content of 50.0%.

AB - Upon release from the stable complex formed with its antitoxin VapB, the toxin VapC (MvpT) of the Gram-negative pathogen Shigella flexneri is capable of globally down-regulating translation by specifically cleaving initiator tRNAfMet in the anticodon region. Recombinant Shigella flexneri VapCD7A harbouring an active-site mutation was overexpressed in Escherichia coli, purified to homogeneity and crystallized by the vapour-diffusion technique. A preliminary X-ray crystallographic analysis shows that the crystals diffracted to at least 1.9 Å resolution at a synchrotron X-ray source and belonged to the trigonal space group in the hexagonal setting, H3, with unit-cell parameters a = b = 120.1, c = 52.5 Å, α = β = 90, γ = 120°. The Matthews coefficient is 2.46 Å3 Da-1, suggesting two molecules per asymmetric unit and corresponding to a solvent content of 50.0%.

U2 - 10.1107/S1744309113014012

DO - 10.1107/S1744309113014012

M3 - Journal article

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SN - 2053-230X

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JO - Acta Crystallographica Section F: Structural Biology Communications

JF - Acta Crystallographica Section F: Structural Biology Communications

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ER -

Protein expression, crystallization and preliminary X-ray crystallographic analysis of the isolated Shigella flexneri VapC toxin

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