Characterization of Acetylcholinesterase in Caco-2 Cells
| Autor: | Kenneth A. Skau, Giovanni M. Pauletti, Lauren R. Plageman |
|---|---|
| Rok vydání: | 2002 |
| Předmět: |
0301 basic medicine
Dopamine General Biochemistry Genetics and Molecular Biology Substrate Specificity Butyrylthiocholine 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Enzyme Stability Humans chemistry.chemical_classification Binding Sites biology Cell Membrane Active site Substrate (chemistry) Cell Differentiation Acetylcholinesterase Acetylcholine Enzyme assay Kinetics 030104 developmental biology Membrane Enzyme Solubility chemistry Biochemistry 030220 oncology & carcinogenesis Acetylthiocholine biology.protein Caco-2 Cells Subcellular Fractions |
| Zdroj: | Experimental Biology and Medicine. 227:480-486 |
| ISSN: | 1535-3699 1535-3702 |
| DOI: | 10.1177/153537020222700712 |
| Popis: | Acetylcholinesterase (acetylcholine hydrolase, EC 3.1.1.7) was solubilized from cultured Caco-2 cells. It was established that this enzyme activity is acetylcholinesterase by substrate specificity (acetylthiocholine, acetyl-β-methylthiocholine>propionylthiocholine>butyrylthiocholine), substrate inhibition, and specificity of inhibitors (BW284c51>iso-OMPA). The acetylcholinesterase activity increased proportional to the degree of differentiation of the cells. Most of the enzyme was membrane bound, requiring detergent for solubilization, and the active site faced the external fluid. Only one peak of activity, which corresponded to a monomeric form, could be detected on linear sucrose density gradients. The sedimentation of this form of the enzyme was shifted depending on whether Triton X-100 or Brij 96 detergent was used. These results indicate that the epithelial-derived Caco-2 cells produce predominantly an amphiphilic, monomeric form of acetylcholinesterase that is bound to the plasma membrane and whose catalytic center faces the extracellular fluid. |
| Databáze: | OpenAIRE |
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