Evidence for phosphorylation and oligomeric assembly of presenilin 1

Proc Natl Acad Sci U S A. 1997 May 13;94(10):5090-4. doi: 10.1073/pnas.94.10.5090.

Abstract

Pathogenic mutations in presenilin 1 (PS1) are associated with approximately 50% of early-onset familial Alzheimer disease. PS1 is endoproteolytically cleaved to yield a 30-kDa N-terminal fragment (NTF) and an 18-kDa C-terminal fragment (CTF). Using COS7 cells transfected with human PS1, we have found that phorbol 12, 13-dibutyrate and forskolin increase the state of phosphorylation of serine residues of the human CTF. Phosphorylation of the human CTF resulted in a shift in electrophoretic mobility from a single major species of 18 kDa to a doublet of 20-23 kDa. This mobility shift was also observed with human PS1 that had been transfected into mouse neuroblastoma (N2a) cells. Treatment of the phosphorylated CTF doublet with phage lambda protein phosphatase eliminated the 20- to 23-kDa doublet while enhancing the 18-kDa species, consistent with the interpretation that the electrophoretic mobility shift was due to the addition of phosphate to the 18-kDa species. The NTF and CTF eluted from a gel filtration column at an estimated mass of over 100 kDa, suggesting that these fragments exist as an oligomerized species. Upon phosphorylation of the PS1 CTF, the apparent mass of the NTF- or CTF-containing oligomers was unchanged. Thus, the association of PS1 fragments may be maintained during cycles of phosphorylation/dephosphorylation of the PS1 CTF.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • COS Cells
  • Chromatography, Gel
  • Electrophoresis, Polyacrylamide Gel
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Macromolecular Substances
  • Membrane Proteins / biosynthesis*
  • Membrane Proteins / chemistry
  • Membrane Proteins / isolation & purification
  • Mice
  • Neuroblastoma
  • PC12 Cells
  • Peptide Fragments / chemistry
  • Peptide Fragments / isolation & purification
  • Phosphoprotein Phosphatases / antagonists & inhibitors
  • Phosphoprotein Phosphatases / metabolism
  • Phosphoproteins / biosynthesis
  • Phosphoproteins / chemistry
  • Phosphoproteins / isolation & purification
  • Phosphorylation
  • Presenilin-1
  • Protein Kinase Inhibitors
  • Protein Kinases / metabolism
  • Rats
  • Recombinant Proteins / biosynthesis
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Serine
  • Transfection
  • Tumor Cells, Cultured

Substances

  • Enzyme Inhibitors
  • Macromolecular Substances
  • Membrane Proteins
  • PSEN1 protein, human
  • Peptide Fragments
  • Phosphoproteins
  • Presenilin-1
  • Protein Kinase Inhibitors
  • Recombinant Proteins
  • Serine
  • Protein Kinases
  • Phosphoprotein Phosphatases