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 Citationer (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 angstrom 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 angstrom, alpha = beta = 90, gamma = 120 degrees. The Matthews coefficient is 2.46 angstrom(3) Da(-1), suggesting two molecules per asymmetric unit and corresponding to a solvent content of 50.0

Originalsprog	Engelsk
Tidsskrift	Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online
Vol/bind	69
Udgave nummer	7
Sider (fra-til)	762-765
Antal sider	4
ISSN	1744-3091
DOI	https://doi.org/10.1107/S1744309113014012
Status	Udgivet - jul. 2013
Udgivet eksternt	Ja

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10.1107/S1744309113014012

Citationsformater

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.
I: Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online, Bind 69, Nr. 7, 07.2013, s. 762-765.

Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › 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, bind 69, nr. 7, s. 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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doi = "10.1107/S1744309113014012",

language = "English",

volume = "69",

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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.

PY - 2013/7

Y1 - 2013/7

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

C2 - 23832203

SN - 1744-3091

VL - 69

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EP - 765

JO - Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online

JF - Acta Crystallographica. Section F: Structural Biology and Crystallization Communications Online

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ER -

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

Abstract

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