A unique hydrophobic domain of rat brain globular acetylcholinesterase for binding to cell membranes |
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Authors: | Christian Andres Mustapha El Mourabit Jean Mark Albert Waksman |
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Institution: | (1) Centre de Neurochimie du C.N.R.S. 5, rue Blaise Pascal, 67084 Strasbourg, France |
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Abstract: | Both salt-soluble and detergent-soluble rat brain globular acetylcholinesterases (SS- and DS- AChE EC 3.1.1.7) are amphiphiles, as shown by detergent dependency of enzymatic activity and binding to liposomes. Proteinase K and papain treatment transformed SS-AChE and DS-AChE into forms that, in absence of detergent, no longer aggregated nor bound to liposomes. In contrast, phosphatidylinositol-specific phospholipase C had no effect on these properties. Labeling DS-AChE with 3-(trifluoromethyl)-3-(m-(125I)-iodophenyl) diazirine (125I]TID) revealed, by polyacrylamide gel electrophoresis under reducing conditions, one single band of 69 kD apparent molecular mass. The same pattern was previously obtained with Bolton and Hunter reagent-labeled enzyme (1). Proteinase K treatment transformed the 11 S 125I]TID labeled AChE into a 4 S form which no longer showed125I-radioactivity and was unable to bind to liposomes. These results are compatible with the existence of a hydrophobic segment present both on salt-soluble and detergent-soluble 11 S AChE as well as on the minor forms 4 S and 7 S. This segment is not linked to the catalytic subunits by disulfide bounds in contrast to the 20 kD non-catalytic subunit described by Inestrosa et al. (2).Abbreviations used AChE
acetylcholinesterase
- SS-AChE
salt-soluble AChE
- DS-AChE
detergent-soluble AChE
- BSA
bovine serum albumin
- ChE
serum (butyryl) cholinesterase
- ConA-Sepharose
concanavalin A-Sepharose 4B
- DMAEBA-Sepharose
dimethylaminoethylbenzoic acid-Sepharose 4B
- PC-Chol-SA liposomes
phosphatidylcholine-cholesterol-stearylamine liposomes
- SDS-PAGE
sodium dodecyl sulfate polyacrylamide gel electrophoresis
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125I-TID
3-(trifluoromethyl)-3-(m-(125I)-iodophenyl) diazirine |
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Keywords: | Acetylcholinesterase globular forms rat brain hydrophobic domain 3-(trifluoromethyl)-3-(m-(125I)-iodophenyl)diazirine liposomes proteinase K |
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