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 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.
authors.affiliation: Karolinska Institutet, Department of Biosciences, Center of Structural Biochemistry, Sweden. wim@csb.ki.se
authors.forename: Winfried
authors.initials: W
authors.lastname: Meining
authors.forename: Johannes
authors.initials: J
authors.lastname: Scheuring
authors.forename: Markus
authors.initials: M
authors.lastname: Fischer
authors.forename: Sevil
authors.initials: S
authors.lastname: Weinkauf
mesh_terms: D000251:Adenosine Triphosphatases / Q000737:chemistry*; D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000737:chemistry*; D003001:Cloning, Molecular; D003460:Crystallization / Q000379:methods; D013293:Enterococcus faecalis / Q000201:enzymology*; D026901:Membrane Transport Proteins / Q000737:chemistry*; D000069816:SEC Translocation Channels; D000081416:SecA Proteins; D012997:Solvents; D014961:X-Ray Diffraction
chemical_list: D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; D000069816:SEC Translocation Channels; D012997:Solvents; D000251:Adenosine Triphosphatases; D000081416:SecA Proteins