p.N17Q/N455Q/N481Q/N486Q/L530X Asn17Gln/Asn455Gln/Asn481Gln/Asn486Gln/Leu530Ter (p.N45Q/N483Q/N509Q/N514Q/L558X Asn45Gln/Asn483Gln/Asn509Gln/Asn514Gln/Leu558Ter in primary sequence with 28 amino-acids signal peptide) N-glycosylation;monomeric and low-glycosylated form of human BChE
Kinetic parameters
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References:
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.