Open AccessCloning, purification, crystallization and preliminary crystallographic analysis of SecA from Enterococcus faecalis
Author(s) -
Meining Winfried,
Scheuring Johannes,
Fischer Markus,
Weinkauf Sevil
Publication year - 2006
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309106017544
Subject(s) - crystallization , enterococcus faecalis , escherichia coli , monoclinic crystal system , cloning (programming) , plasmid , crystallography , chemistry , materials science , gene , biochemistry , crystal structure , organic chemistry , computer science , programming language
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 C 2, 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.