title: Cloning, expression, purification and crystallization of an endotoxin-biosynthesis enzyme from Neisseria meningitidis.
abstract: 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 P2(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.
authors.affiliation: School of Chemistry and Biochemistry, University of Western Australia, 35 Stirling Highway, Crawley, Western Australia 6009, Australia.
authors.forename: Anandhi
authors.initials: A
authors.lastname: Anandan
authors.forename: Susannah
authors.initials: S
authors.lastname: Piek
authors.forename: Charlene M
authors.initials: CM
authors.lastname: Kahler
authors.forename: Alice
authors.initials: A
authors.lastname: Vrielink
mesh_terms: D001425:Bacterial Outer Membrane Proteins / Q000737:chemistry* / Q000378:metabolism; D003001:Cloning, Molecular; D003460:Crystallization; D018360:Crystallography, X-Ray; D004731:Endotoxins / Q000096:biosynthesis*; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009345:Neisseria meningitidis / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism
chemical_list: D001425:Bacterial Outer Membrane Proteins; D004731:Endotoxins