Modulation of the distribution of acetylcholinesterase molecular forms in a murine neuroblastoma x sympathetic ganglion cell hybrid cell line.
Studies were carried out on the polymorphism of acetylcholinesterase (AChE, EC 3.1.1.7) in a neuroblastoma x sympathetic ganglion cell hybrid cell line (T28) and its parental clone (N18TG2). These cells contain the tetrameric (G4, 10S), dimeric (G2, 6.5S) and monomeric (G1, 4S) forms of AchE, but not the collagen-tailed A12(16S) form of the sympathetic ganglion. Three variants of these forms could be distinguished on the basis of their solubility properties: (i) secreted forms which do not interact with the detergent Triton X-100; (ii) cellular forms which may be solubilized in detergent-free buffer and which interact reversibly with Triton X-100; (iii) cellular forms which require detergent for solubility, and aggregate in its absence. By using a nonpenetrating inhibitor, we demonstrated that, in T28 stationary cells, the cellular G4 form is associated with the plasma membrane, whereas the G1 form is intracellular. During induction of AChE activity in T28 cells, the relative proportion of the G4 form increases, suggesting, in agreement with previous observations, that G1 is a metabolic precursor of G4. The evolution of AChE molecular forms released into the culture medium closely resembles that of the cellular forms. The preferential accumulation of the G4 molecules does not simply depend on the cellular level of G1. It is favoured by culture conditions which promote morphological differentiation, but does not require the actual extension of neurites. T28 cells as well as other neuroblastoma-derived cells appear to be useful experimental materials to investigate the regulatory mechanisms underlying the maturation of AChE globular forms.
D000110:Acetylcholinesterase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D002460:Cell Line; D002999:Clone Cells; D003470:Culture Media; D004790:Enzyme Induction; D005728:Ganglia, Sympathetic / Q000201:enzymology; D006822:Hybrid Cells / Q000201:enzymology; D046911:Macromolecular Substances; D009447:Neuroblastoma / Q000201:enzymology*; D011110:Polymorphism, Genetic
D003470:Culture Media; D046911:Macromolecular Substances; D000110:Acetylcholinesterase