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

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2006 Jun 1;62(Pt 6):583-5. doi: 10.1107/S1744309106017544. Epub 2006 May 31.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphatases / chemistry*
  • Amino Acid Substitution
  • Bacterial Proteins / chemistry*
  • Cloning, Molecular
  • Crystallization / methods
  • Enterococcus faecalis / enzymology*
  • Membrane Transport Proteins / chemistry*
  • SEC Translocation Channels
  • SecA Proteins
  • Solvents
  • X-Ray Diffraction

Substances

  • Bacterial Proteins
  • Membrane Transport Proteins
  • SEC Translocation Channels
  • Solvents
  • Adenosine Triphosphatases
  • SecA Proteins

Associated data

  • PDB/1M6N
  • PDB/1NKT