An asymmetric form of muscle acetylcholinesterase contains three subunit types and two enzymic activities in one molecule.
We have purified completely the principal asymmetric ("heavy") form of acetylcholinesterase (Ac-ChoEase; EC 3.1.1.7) from chick muscle (i.e., the synaptic form in the twitch muscle fibers) by using a monoclonal antibody that recognizes AcChoEase but not pseudocholinesterase (ChoEase; cholinesterase, EC 3.1.1.8). The purified protein exhibits catalytic and inhibition properties characteristic of AcChoEase and ChoEase and contains three distinct subunits of apparent sizes 110 kDa, 72 kDa, and 58 kDa in the ratio 2:2:1. The discovery of an AcChoEase/ChoEase hybrid asymmetric form has been further supported by (i) the identification of active site properties of AcChoEase in the 110-kDa subunit and of ChoEase in the 72-kDa subunit, (ii) the purification or precipitation of both activities together by, also, a ChoEase-specific monoclonal antibody, and (iii) evidence that all subunits are bound in the asymmetric forms by disulfide bonds. The 58-kDa subunit is the only one that is sensitive to digestion with purified collagenase; it carries the collagenous "tail" of the asymmetric form. A model is proposed for this form of AcChoEase.
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Medical Research Council Molecular Neurobiology Unit, Medical Research Council Centre, Cambridge, England.
Randall
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WR
Barnard
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EA
D000110:Acetylcholinesterase / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002091:Butyrylcholinesterase / Q000302:isolation & purification* / Q000378:metabolism; D002645:Chickens; D002802:Cholinesterases / Q000302:isolation & purification*; D009132:Muscles / Q000201:enzymology* / Q000276:immunology; D011487:Protein Conformation
D000911:Antibodies, Monoclonal; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase; D002802:Cholinesterases