Butyrylcholinesterase from chicken brain is smaller than that from serum: its purification, glycosylation, and membrane association.
Applying a new four-step isolation procedure, we have purified butyrylcholinesterase (BChE) from chicken serum to homogeneity with more than 250 U/mg specific activity. The serum enzyme was used for producing monoclonal antibodies. These BChE-specific also recognize BChE from brain, and thus enabled us to isolate the enzymes from embryonic and adult brain that occur only in minute amounts. More than 50% of the brain BChE is membrane-bound. The catalytic and inhibition properties of brain BChE are similar to those of serum BChE. However on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the serum enzyme is represented by a double-band of 79/82 kDa, whereas the brain enzyme has a size of 74 kDa. Limited digestion of the serum and brain preparations by V8-protease leads to similar peptide patterns. Enzymatic deglycosylation shows that their core proteins consist of 59-kDa subunits and that the different molecular weights are due to different glycosylation patterns. The differently sized glycosylation parts of brain and serum BChE may indicate that they subserve different functions. Furthermore, the membrane-bound brain BChE can be solubilized by Pronase or protease K, but not by phosphatidylinositol-specific phospholipase C.
Treskatis
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S
Max-Planck-Institut für Entwicklungsbiologie, Tübingen, F.R.G.
Ebert
C
C
Layer
P G
PG
D000818:Animals; D001921:Brain / Q000201:enzymology* / Q000378:metabolism / Q000648:ultrastructure; D002091:Butyrylcholinesterase / Q000097:blood* / Q000737:chemistry* / Q000302:isolation & purification; D002462:Cell Membrane / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure; D002499:Centrifugation, Density Gradient; D002645:Chickens; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D019286:Endopeptidase K; D006031:Glycosylation; D008970:Molecular Weight; D010449:Peptide Mapping; D011402:Pronase / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000494:pharmacology
D002091:Butyrylcholinesterase; D012697:Serine Endopeptidases; D019286:Endopeptidase K; D011402:Pronase