Open AccessCloning, expression, purification and crystallization of an endotoxin‐biosynthesis enzyme from Neisseria meningitidis
Author(s) -
Anandan Anandhi,
Piek Susannah,
Kahler Charlene M.,
Vrielink Alice
Publication year - 2012
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309112042236
Subject(s) - periplasmic space , escherichia coli , enzyme , neisseria meningitidis , biochemistry , crystallization , transmembrane domain , chemistry , cloning (programming) , transferase , bacteria , biology , amino acid , gene , genetics , organic chemistry , computer science , programming language
The enzyme phosphoethanolamine transferase A is involved in the addition of phosphoethanolamine moieties to lipid A in Neisseria meningitidis . The enzyme is composed of an N‐terminal transmembrane domain and a C‐terminal soluble domain that is present in the periplasm of the bacteria. A membrane‐deletion construct of the enzyme was designed and expressed in Escherichia coli . Well ordered crystals that diffracted to 1.7 Å resolution were obtained by carrying out a limited trypsin digestion of the protein to remove a predicted N‐terminal disordered portion. The crystals belonged to space group P 2 1 , with unit‐cell parameters a = 44.3, b = 71.6, c = 49.9 Å, β = 109.2°, and contained one molecule in the asymmetric unit.