Analysis of presenilin 1 and presenilin 2 expression and processing by newly developed monoclonal antibodies.
Because distinct mutations in presenilin 1 and presenilin 2 are a major cause of early-onset familial Alzheimer's disease, we generated four monoclonal antibodies for the identification, localization, and investigation of presenilins in various cell lines and tissues from patients and controls. We show that these antibodies are specific for the N- and C-terminal domains of human presenilin 1 and presenilin 2. They recognize presenilin full-length proteins and their approximately 28-35 kDa N-terminal fragments and approximately 18-20 kDa C-terminal fragments. None of the antibodies showed cross-reaction in their specific detection ability. We demonstrated that presenilin 1 and presenilin 2 are proteolytically processed in human glioma cell lines, transfected and untransfected human neuroblastoma SH-SY5Y cells, COS-7 cells, rat cerebellar neuronal ST15 cells, mouse and human brain. Remarkably, we observed that presenilin 2 is alternatively cleaved during apoptosis, producing smaller C-terminal fragments. By analyzing the subcellular distribution of presenilins, we found reticular and fine vesicular staining throughout the cell bodies. In addition, staining of Golgi compartments and the perinuclear envelope was observed. Alzheimer's disease brain showed strong immunoreactivity of presenilin 1 in reactive astrocytes and senile plaques. This high expression of presenilin 1 may explain the increased production and accumulation of the amyloid-beta peptide in patients with sporadic Alzheimer's disease in the absence of familial presenilin mutation.
Diehlmann
A
A
ZMBH, Heidelberg, Germany. anke.d@zmbh.uni-heidelberg.de
Ida
N
N
Weggen
S
S
Grünberg
J
J
Haass
C
C
Masters
C L
CL
Bayer
T A
TA
Beyreuther
K
K
D000544:Alzheimer Disease / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis / Q000276:immunology* / Q000302:isolation & purification; D000918:Antibody Specificity; D017209:Apoptosis; D001253:Astrocytes / Q000378:metabolism / Q000473:pathology; D001921:Brain / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D002460:Cell Line; D003429:Cross Reactions / Q000276:immunology; D006801:Humans; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D051379:Mice; D008970:Molecular Weight; D009154:Mutation; D009474:Neurons / Q000378:metabolism / Q000473:pathology; D015388:Organelles / Q000378:metabolism; D010446:Peptide Fragments / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D058225:Plaque, Amyloid / Q000378:metabolism; D053764:Presenilin-1; D053766:Presenilin-2; D011499:Protein Processing, Post-Translational; D051381:Rats; D014162:Transfection
D000911:Antibodies, Monoclonal; D008565:Membrane Proteins; C508853:PSEN1 protein, human; C508861:PSEN2 protein, human; D010446:Peptide Fragments; D053764:Presenilin-1; D053766:Presenilin-2