Pro-apoptotic protein–protein interactions of the extended N-AChE terminus

Abstract

The N-terminally extended ''synaptic'' acetylcholinesterase variant N-AChE-S operates to promote apoptosis; however, the protein partners involved in this function remain unknown. Here, we report that when microinjected to fertilized mouse oocytes, N-AChE-S caused embryonic death as early as the zygotic stage. To identify the putative protein partners involved, we first tried yeast two hybrid screening, but this approach failed, probably because of the N-AChE-S-induced lethality. In contrast, sequence analysis and a corresponding peptide array revealed possible partners, which were validated by co-immunoprecipitation. These include the kinases GSK3, Aurora and GAK, the membrane integrin receptors, and the death receptor FAS. Each of these could potentially modulate N-AChE-S-induced apoptosis with possible therapeutic value for the treatment of Alzheimer's disease.

Acetylcholinesterase is associated with apoptosis in β cells and contributes to insulin-dependent diabetes mellitus pathogenesis

“…Previously, we reported that AChE was expressed during apoptosis induced by various stimuli in a number of cell lines [4]. AChE has been proposed to play a pivotal role in apoptosome formation [5,6], and synaptic AChE with an extended N-terminus induces apoptosis in neurons by interacting with other proteins [7,8]. Moreover, the appearance of this enzyme is considered as a marker for apoptosis under certain circumstances [9][10][11].…”

Acetylcholinesterase Variants in Alzheimer’s Disease: From Neuroprotection to Programmed Cell Death

“…Recombinant N-AChE-S, but none of the other variants, induced programmed cell death, both in microinjected early mouse embryos and in transfected cultured cells of different species and embryonic origins [9,10] ( fig. 1 a, b).…”

“…Brain tissue samples from AD patients and matched nondemented controls as well as transgenic mice overexpressing specific human AChE variants, the doubly mutated amyloid APPswe 'Swedish' gene or both were used in this study. Fluorescence in situ hybridization (FISH) and immunohistochemistry were used to label mRNA and protein products, respectively [10,11] . siRNA agents were used to suppress AChE expression in cultured cells and in vivo, and highly purified AChE-R produced in transgenic plants [12] enabled both biochemical and cellular loss and gain of function tests of these variants ex vivo and in vivo.…”

“…Intriguingly, the AChE gene can form multiple 3 -alternatively spliced variants [5,6] as well as alternate 5 -transcripts and N-terminally extended protein products [7][8][9][10] . All of these AChE variants share similar hydrolytic activity, but their subcellular localization, cellular properties and physiological roles were found to be different and sometimes inverse to each other, as is often the case with alternative splicing products of single genes [11] .…”

The Amyloid Precursor Protein Represses Expression of Acetylcholinesterase in Neuronal Cell Lines

“…AChE and APP both have roles in cell adhesion and synaptic integrity (3,77), so perhaps this novel relationship serves to regulate these non-catalytic functions. Finally, AChE has been shown to have a role in apoptosis, being up-regulated by certain apoptotic stimuli and then participating in the process of apoptosis (78,79), including in AD (80). As such, this down-regulation of AChE might be an example of a novel neuroprotective function of the APP holoprotein.…”