Monoclonal antibodies allow precipitation of esterasic but not peptidasic activities associated with butyrylcholinesterase.
Commercially available and affinity-purified butyrylcholinesterases isolated from human serum were examined for their esterasic activity and their ability to hydrolyze various neuropeptides, including neurotensin, substance P, and leucine-enkephalin. The three pools that displayed the lowest esterasic activities were shown to hydrolyze neurotensin with the same HPLC degradative pattern. By contrast, noticeable qualitative and quantitative discrepancies were observed when hydrolyses of substance P and leucine-enkephalin by these three butyrylcholinesterase pools were studied. The pool that exhibited the highest esterasic activity appeared to be homogeneously constituted by 90- and 180-kDa protein bands by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis and was totally unable to hydrolyze these three neuropeptides. This suggested that the three other butyrylcholinesterase preparations could be contaminated by exogenous peptidases. This was confirmed by means of three distinct monoclonal antibodies directed toward human serum butyrylcholinesterase. The three IgG-purified fractions precipitated the esterasic activity, whereas they failed to precipitate the neuropeptide-hydrolyzing activities whatever the substrate examined. Altogether, these results demonstrate that peptidases associated with butyrylcholinesterase are contaminating enzymes that cannot be considered as intrinsic activities of this enzyme.
Checler
F
F
Institut de Pharmacologie Moléculaire et Cellulaire du CNRS, Université de Nice-Sophia Antipolis, Valbonne, France.
Grassi
J
J
Masson
P
P
Vincent
J P
JP
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D002091:Butyrylcholinesterase / Q000097:blood*; D002802:Cholinesterases / Q000097:blood*; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000097:blood*; D004950:Esterases / Q000097:blood*; D006801:Humans; D006868:Hydrolysis; D007163:Immunosorbent Techniques; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009496:Neurotensin / Q000378:metabolism; D009842:Oligopeptides / Q000378:metabolism; D013373:Substance P / Q000378:metabolism; D013379:Substrate Specificity
D000911:Antibodies, Monoclonal; D009842:Oligopeptides; D013373:Substance P; D009496:Neurotensin; D004950:Esterases; D002091:Butyrylcholinesterase; D002802:Cholinesterases; D010450:Endopeptidases