Purification and properties of the membrane-bound acetylcholinesterase from adult rat brain
| Autor: | J.P. Zanetta, Z. Rakonczay, H. Schenk, Guy Vincendon, J. Mallol |
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| Rok vydání: | 1981 |
| Předmět: |
Octoxynol
Size-exclusion chromatography Chromatography Affinity Chromatography DEAE-Cellulose Polyethylene Glycols chemistry.chemical_compound Affinity chromatography Centrifugation Density Gradient Animals Sodium dodecyl sulfate Polyacrylamide gel electrophoresis chemistry.chemical_classification Chromatography Isoelectric focusing Cell Membrane Brain General Medicine Chromatography Agarose Acetylcholinesterase Rats Molecular Weight Electrophoresis Enzyme Biochemistry chemistry Chromatography Gel Electrophoresis Polyacrylamide Gel Isoelectric Focusing |
| Zdroj: | Biochimica et Biophysica Acta (BBA) - Enzymology. 657:243-256 |
| ISSN: | 0005-2744 |
| DOI: | 10.1016/0005-2744(81)90148-0 |
| Popis: | The membrane-bound acetylcholinesterase (acetylcholine acetylhydrolase, EC 3.1.1.7) from adult rat brain has been purified to homogeneity using sequential affinity chromatography on Con A-Sepharose and on dimethyl-aminoethylbenzoic acid-Sepharose 4B followed by DEAE-cellulose chromatography. The yield of the purified enzyme (specific activity: 3068 U/mg protein) is higher than 50%. Polyacrylamide gel electrophoresis in the presence of Triton X-100 gives only one band with acetylcholinesterase activity. With the exception of electrofocusing and pore gradient electrophoresis, where a multiple band pattern was detected (which seems to be artefactual), the enzyme appears to be homogenous. Gel filtration and sucrose density gradient centrifugation in the presence of Triton X-100 give only one symmetrical peak, with a calculated molecular weight of 328 000. Since polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) and mercaptoethanol gives only one band with a molecular weight of 74 500, a tetrametric structure can be postulated for the membrane-bound acetylcholinesterase from rat brain. |
| Databáze: | OpenAIRE |
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