Abstract
Background
Alzheimer’s disease (AD) is a progressive neurodegenerative disorder characterized by the deposition of extracellular senile plaques composed of amyloid β-peptide (Aβ). Whereas most cases of AD occur sporadically, about 10% of AD cases are inherited as a fully penetrant autosomal dominant trait. Mutations in the recently cloned Presenilin genes (PS-1 and PS-2) are by far the most common cause of early onset familial AD.
Materials and Methods
Cellular expression of endogenous and overexpressed PS proteins was analyzed by immunocytochemistry and metabolic labeling followed by immunoprecipitation. In vivo phosphorylation sites of PS proteins were analyzed by extensive mutagenesis.
Results
PS-1 as well as PS-2 proteins were localized predominantly within the endoplasmic reticulum (ER). However, small amounts of the PS proteins were detected within the Golgi compartment, where they colocalize with the β-amyloid precursor protein (βAPP). The PS-2 protein was found to be highly phosphorylated. whereas very little phosphorylation was observed for PS-1. The selective phosphorylation of PS-2 occurs exclusively on serine residues. In vivo phosphorylation of PS-2 was mapped to serine residues 7, 9, and 19 within an acidic stretch at the N terminus, which is absent in PS-1. Casein kinase (CK)-l and CK-2 were shown to phosphorylate the N terminus of PS-2 in vitro.
Conclusions
The majority of PS proteins were detected in the ER where little if any proteolytic processing of βAPP was reported. ER retention of PS proteins might occur by intramolecular aggregation. Small amounts of PS proteins were also detected in the Golgi where they colocalized with βAPP. This might suggest that potential interactions between PS proteins and βAPP could occur within the Golgi. Selective phosphorylation of PS-2 proteins within the acidic domain missing in PS-1 indicates differences in the biological functions and regulation of the two highly homologous proteins.
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Acknowledgments
This work was supported by grants from the Deutsche Forschungsgemeinschaft (SFB317HA 1737/2-1; MU 467/8-1), by the Boehringer Ingelheim KG. (to CH), by National Institutes of Health Grants AG 06173 and AG 12749 (to DJS), and by the Alzheimer Association of Ontario, the Medical Research Council of Canada and the Canadian Genetic Disease Network (to PSH). We thank Dr. Walter Pyerin for providing human recombinant CK-2, Dr. Edward Koo for providing monoclonal antibodies to βAPP, Dr. Dale Schenk for providing antibody B5, and Dr. Martin Citron for critical comments and discussion.
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Walter, J., Capell, A., Grünberg, J. et al. The Alzheimer’s Disease-Associated Presenilins Are Differentially Phosphorylated Proteins Located Predominantly within the Endoplasmic Reticulum. Mol Med 2, 673–691 (1996). https://doi.org/10.1007/BF03401652
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DOI: https://doi.org/10.1007/BF03401652