TY - JOUR
T1 - Cloning, expression, purification and crystallization of an endotoxin-biosynthesis enzyme from Neisseria meningitidis
AU - Anandan, Anandhi
AU - Piek, Susannah
AU - Kahler, Charlene
AU - Vrielink, Alice
PY - 2012/12/1
Y1 - 2012/12/1
N2 - 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 P21, with unit-cell parameters a = 44.3, b = 71.6, c = 49.9 Å, [beta] = 109.2°, and contained one molecule in the asymmetric unit.
AB - 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 P21, with unit-cell parameters a = 44.3, b = 71.6, c = 49.9 Å, [beta] = 109.2°, and contained one molecule in the asymmetric unit.
U2 - 10.1107/S1744309112042236
DO - 10.1107/S1744309112042236
M3 - Article
SN - 1744-3091
VL - 68
SP - 1494
EP - 1497
JO - ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS
JF - ACTA CRYSTALLOGRAPHICA SECTION F-STRUCTURAL BIOLOGY AND CRYSTALLIZATION COMMUNICATIONS
IS - 12
ER -