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. 2006 Jun 1;62(Pt 6):583-5.
doi: 10.1107/S1744309106017544. Epub 2006 May 31.

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

Winfried Meining  1 , Johannes Scheuring, Markus Fischer, Sevil Weinkauf
Affiliations

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

Winfried Meining et al. Acta Crystallogr Sect F Struct Biol Cryst Commun. .

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.

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Figures

Figure 1
Figure 1
Crystal of native SecA from E. faecalis. The scale bar corresponds to 0.1 mm. See text for a description of crystallization conditions.
See this image and copyright information in PMC

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