Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis

Winfried Meining; Johannes Scheuring; Markus Fischer; Sevil Weinkauf

doi:10.1107/S1744309106017544

Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis

Winfried Meining
, Johannes Scheuring
, Markus Fischer
, Sevil Weinkauf

Associate Professorship of Electron Microscopy

Karolinska Institutet
Technical University of Munich

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

The gene coding for SecA from Enterococcus faecalis was cloned and overexpressed in Escherichia coli. In this protein, the lysine at position 6 was replaced by an asparagine in order to reduce sensitivity towards proteases. The modified protein was purified and crystallized. Crystals diffracting to 2.4 Å resolution were obtained using the vapour-diffusion technique. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 203.4, b = 49.8, c = 100.8 Å, α = γ = 90.0, β = 119.1°. A selenomethionine derivative was prepared and is currently being tested in crystallization trials.

Original language	English
Pages (from-to)	583-585
Number of pages	3
Journal	Acta Crystallographica Section F: Structural Biology and Crystallization Communications
Volume	62
Issue number	6
DOIs	https://doi.org/10.1107/S1744309106017544
State	Published - Jun 2006

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10.1107/S1744309106017544

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title = "Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis",

abstract = "The gene coding for SecA from Enterococcus faecalis was cloned and overexpressed in Escherichia coli. In this protein, the lysine at position 6 was replaced by an asparagine in order to reduce sensitivity towards proteases. The modified protein was purified and crystallized. Crystals diffracting to 2.4 {\AA} resolution were obtained using the vapour-diffusion technique. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 203.4, b = 49.8, c = 100.8 {\AA}, α = γ = 90.0, β = 119.1°. A selenomethionine derivative was prepared and is currently being tested in crystallization trials.",

author = "Winfried Meining and Johannes Scheuring and Markus Fischer and Sevil Weinkauf",

year = "2006",

month = jun,

doi = "10.1107/S1744309106017544",

language = "English",

volume = "62",

pages = "583--585",

journal = "Acta Crystallographica Section F: Structural Biology and Crystallization Communications",

issn = "1744-3091",

publisher = "International Union of Crystallography",

number = "6",

}

Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis. / Meining, Winfried; Scheuring, Johannes; Fischer, Markus et al.
In: Acta Crystallographica Section F: Structural Biology and Crystallization Communications, Vol. 62, No. 6, 06.2006, p. 583-585.

Research output: Contribution to journal › Article › peer-review

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T1 - Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis

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AU - Scheuring, Johannes

AU - Fischer, Markus

AU - Weinkauf, Sevil

PY - 2006/6

Y1 - 2006/6

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AB - The gene coding for SecA from Enterococcus faecalis was cloned and overexpressed in Escherichia coli. In this protein, the lysine at position 6 was replaced by an asparagine in order to reduce sensitivity towards proteases. The modified protein was purified and crystallized. Crystals diffracting to 2.4 Å resolution were obtained using the vapour-diffusion technique. The crystals belong to the monoclinic space group C2, with unit-cell parameters a = 203.4, b = 49.8, c = 100.8 Å, α = γ = 90.0, β = 119.1°. A selenomethionine derivative was prepared and is currently being tested in crystallization trials.

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DO - 10.1107/S1744309106017544

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

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

IS - 6

ER -

Cloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis

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