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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 A 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 A, alpha = gamma = 90.0, beta = 119.1 degrees. A selenomethionine derivative was prepared and is currently being tested in crystallization trials.

SUBMITTER: Meining W

PROVIDER: S-EPMC2243102 | biostudies-literature | 2006 Jun

REPOSITORIES: biostudies-literature

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

Meining Winfried W Scheuring Johannes J Fischer Markus M Weinkauf Sevil S

Acta crystallographica. Section F, Structural biology and crystallization communications 20060531 Pt 6

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 A 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 A, alpha = gamma = 9 ...[more]

PMID: 16754988

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