Glycoproteomic characterization of butyrylcholinesterase from human plasma.
Human butyrylcholinesterase (hBChE) is a highly glycosylated protein present in human plasma. The enzyme hydrolyses choline esters, for example benzoylcholine, butyrylthiocholine and acetylthiocholine as well as noncholine esters like heroin and aspirin. hBChE is primarily involved in neuronal transmission and is a potential bioscavenger of toxic organophosphates to protect acetylcholinesterase. A prerequisite for the therapeutic use of hBChE is a detailed characterization of this glycoprotein purified from human plasma. In this study, MS/MS could confirm most of the protein backbone, including the N- and the C-terminus. Site-specific analysis of all nine potential N-glycosylation sites revealed mainly mono- and disialylated N-glycans to be present on this glycoprotein. Sialic acids (Neu5Ac) are mainly alpha2,6-linked, however a fraction of the N-glycans contained Neu5Ac also in alpha2,3 linkage. On monosialylated N-glycans, sialic acid is exclusively located on the 3-arm and in alpha2,6 linkage, as verified by 2D-HPLC and exoglycosidase digests of 2-aminopyridine (PA)-labelled N-glycans. This first comprehensive glycoproteomic analysis of the important human plasma glycoprotein BChE did not give any indication of O-glycosylation or any other kind of PTMs as previously postulated.
Kolarich
Daniel
D
Biochemistry Division, Department of Chemistry, University of Natural Resources and Applied Life Sciences, Vienna, Austria. daniel.kolarich@mq.edu.au
Weber
Alfred
A
Pabst
Martin
M
Stadlmann
Johannes
J
Teschner
Wolfgang
W
Ehrlich
Hartmut
H
Schwarz
Hans-Peter
HP
Altmann
Friedrich
F
D000595:Amino Acid Sequence; D002091:Butyrylcholinesterase / Q000097:blood*; D002240:Carbohydrate Sequence; D002853:Chromatography, Liquid; D006023:Glycoproteins / Q000737:chemistry*; D006026:Glycoside Hydrolases / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D025942:Oligosaccharides, Branched-Chain; D011134:Polysaccharides / Q000737:chemistry; D040901:Proteomics / Q000379:methods*
D006023:Glycoproteins; D025942:Oligosaccharides, Branched-Chain; D011134:Polysaccharides; D002091:Butyrylcholinesterase; D006026:Glycoside Hydrolases