title: The Alzheimer's disease-associated presenilins are differentially phosphorylated proteins located predominantly within the endoplasmic reticulum.
abstract: BACKGROUND
  Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by the deposition of extracellular senile plaques composed of amyloid beta-peptide (A beta). 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.
  
  
  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 beta-amyloid precursor protein (beta APP). 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)-1 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 beta APP 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 beta APP. This might suggest that potential interactions between PS proteins and beta APP 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.
authors.affiliation: Central Institute of Mental Health, Department of Molecular Biology, Mannheim, Germany.
authors.forename: J
authors.initials: J
authors.lastname: Walter
authors.forename: A
authors.initials: A
authors.lastname: Capell
authors.forename: J
authors.initials: J
authors.lastname: Grünberg
authors.forename: B
authors.initials: B
authors.lastname: Pesold
authors.forename: A
authors.initials: A
authors.lastname: Schindzielorz
authors.forename: R
authors.initials: R
authors.lastname: Prior
authors.forename: M B
authors.initials: MB
authors.lastname: Podlisny
authors.forename: P
authors.initials: P
authors.lastname: Fraser
authors.forename: P S
authors.initials: PS
authors.lastname: Hyslop
authors.forename: D J
authors.initials: DJ
authors.lastname: Selkoe
authors.forename: C
authors.initials: C
authors.lastname: Haass
mesh_terms: D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism*; D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism; D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D017931:DNA Primers; D004721:Endoplasmic Reticulum / Q000737:chemistry; D005786:Gene Expression Regulation / Q000235:genetics; D006056:Golgi Apparatus / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed / Q000235:genetics; D010766:Phosphorylation; D053764:Presenilin-1; D053766:Presenilin-2; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000235:genetics; D016415:Sequence Alignment; D014162:Transfection / Q000235:genetics
chemical_list: D016564:Amyloid beta-Protein Precursor; D017931:DNA Primers; D008565:Membrane Proteins; C508853:PSEN1 protein, human; C508861:PSEN2 protein, human; D053764:Presenilin-1; D053766:Presenilin-2; D011493:Protein Kinase C