Enhanced production and oligomerization of the 42-residue amyloid beta-protein by Chinese hamster ovary cells stably expressing mutant presenilins.
Mutations in the presenilin 1 (PS1) and presenilin 2 (PS2) genes cause the most common and aggressive form of early onset familial Alzheimer's disease. To elucidate their pathogenic mechanism, wild-type (wt) or mutant (M146L, C410Y) PS1 and wt or mutant (M239V) PS2 genes were stably transfected into Chinese hamster ovary cells that overexpress the beta-amyloid precursor protein (APP). The identity of the 43-45-kDa PS1 holoproteins was confirmed by N-terminal radiosequencing. PS1 was rapidly processed (t1/2 = 40 min) in the endoplasmic reticulum into stable fragments. Wild-type and mutant PS2 holoproteins exhibited similar half lives (1.5 h); however, their endoproteolytic fragments showed both mutation-specific and cell type-specific differences. Mutant PS1 or PS2 consistently induced a 1.4-2.5-fold increase (p < 0.001) in the relative production of the highly amyloidogenic 42-residue form of amyloid beta-protein (Abeta42) as determined by quantitative immunoprecipitation and by enzyme-linked immunosorbent assay. In mutant PS1 and PS2 cell lines with high increases in Abeta42/Abetatotal ratios, spontaneous formation of low molecular weight oligomers of Abeta42 was observed in media, suggesting enhanced Abeta aggregation from the elevation of Abeta42. We conclude that mutant PS1 and PS2 proteins enhance the proteolysis of beta-amyloid precursor protein by the gamma-secretase cleaving at Abeta residue 42, thereby promoting amyloidogenesis.
Xia
W
W
Department of Neurology, Harvard Medical School and Center for Neurologic Diseases, Brigham and Women's Hospital, Boston, Massachusetts 02115, USA.
Zhang
J
J
Kholodenko
D
D
Citron
M
M
Podlisny
M B
MB
Teplow
D B
DB
Haass
C
C
Seubert
P
P
Koo
E H
EH
Selkoe
D J
DJ
D016229:Amyloid beta-Peptides / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D000818:Animals; D001704:Biopolymers; D016466:CHO Cells; D006224:Cricetinae; D003412:Cricetulus; D006868:Hydrolysis; D008565:Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D053764:Presenilin-1; D053766:Presenilin-2; D014162:Transfection
D016229:Amyloid beta-Peptides; D001704:Biopolymers; D008565:Membrane Proteins; D053764:Presenilin-1; D053766:Presenilin-2