Paper Report for: Nachon_2002_Eur.J.Biochem_269_630
Reference
Title: Engineering of a monomeric and low-glycosylated form of human butyrylcholinesterase: expression, purification, characterization and crystallization Nachon F, Nicolet Y, Viguie N, Masson P, Fontecilla-Camps JC, Lockridge O Ref: European Journal of Biochemistry, 269:630, 2002 : PubMed
Human butyrylcholinesterase (BChE; EC 3.1.1.8) is of particular interest because it hydrolyzes or scavenges a wide range of toxic compounds including cocaine, organophosphorus pesticides and nerve agents. The relative contribution of each N-linked glycan for the solubility, the stability and the secretion of the enzyme was investigated. A recombinant monomeric BChE lacking four out of nine N-glycosylation sites and the C-terminal oligomerization domain was stably expressed as a monomer in CHO cells. The purified recombinant BChE showed catalytic properties similar to those of the native enzyme. Tetragonal crystals suitable for X-ray crystallography studies were obtained; they were improved by recrystallization and found to diffract to 2.0 A resolution using synchrotron radiation. The crystals belong to the tetragonal space group I422 with unit cell dimensions a = b = 154.7 A, c = 124.9 A, giving a Vm of 2.73 A3 per Da (estimated 60% solvent) for a single molecule of recombinant BChE in the asymmetric unit. The crystal structure of butyrylcholinesterase will help elucidate unsolved issues concerning cholinesterase mechanisms in general.
Nachon F, Nicolet Y, Viguie N, Masson P, Fontecilla-Camps JC, Lockridge O (2002) Engineering of a monomeric and low-glycosylated form of human butyrylcholinesterase: expression, purification, characterization and crystallization European Journal of Biochemistry269: 630-7
Nachon F, Nicolet Y, Viguie N, Masson P, Fontecilla-Camps JC, Lockridge O (2002) European Journal of Biochemistry269: 630-7
