Acetylcholinesterase in mouse neuroblastoma NB2A cells: analysis of production, secretion, and molecular forms.
The mouse neuroblastoma cell line NB2A produces cellular and secreted acetylcholinesterase (AChE). After incubation of the cells for 4 days the ratio between AChE secreted into the medium and AChE in the cells was 1:1. The cell-associated enzyme could be subdivided into soluble AChE (25%) and detergent-soluble AChE (75%). Both extracts contained predominantly monomeric AChE (4.6S) and minor amounts of tetrameric AChE (10.6S), whereas the secreted AChE in the culture supernatant contained only the tetrameric form. All forms were partially purified by affinity chromatography. It could be demonstrated that the secretory and the intracellular soluble tetramers were hydrophilic, whereas the detergent-soluble tetramer was an amphiphilic protein. On the other hand the soluble and the detergent-soluble monomeric forms were amphiphilic and their activity depended on the presence of detergent. By digestion with proteinase K amphiphilic monomeric and tetrameric AChE could be converted to a hydrophilic form that no longer required detergent for catalytic activity. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of [3H]diisopropylfluorophosphate-labelled AChE gave one band at 64 kilodaltons (kD) under reducing conditions and two additional bands at 120 kD and 140 kD under nonreducing conditions.
Stieger
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S
Institut für Biochemie und Molekularbiologie, Universität Bern, Switzerland.
Bütikofer
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P
Wiesmann
U N
UN
Brodbeck
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U
D000110:Acetylcholinesterase / Q000096:biosynthesis / Q000378:metabolism*; D000818:Animals; D055598:Chemical Phenomena; D002621:Chemistry; D019286:Endopeptidase K; D008968:Molecular Conformation; D009447:Neuroblastoma / Q000378:metabolism* / Q000473:pathology; D012697:Serine Endopeptidases / Q000494:pharmacology; D014407:Tumor Cells, Cultured; D010738:Type C Phospholipases / Q000494:pharmacology
D000110:Acetylcholinesterase; D010738:Type C Phospholipases; D012697:Serine Endopeptidases; D019286:Endopeptidase K