The Alzheimer amyloid precursor protein. Identification of a stable intermediate in the biosynthetic/degradative pathway.
The amyloid forming beta-peptide of Alzheimer's disease is synthesized as part of a larger integral membrane precursor protein (beta APP) of which three alternatively spliced versions of 695, 751, and 770 amino acids have been described. A fourth beta APP form of 563 amino acids does not contain the beta-peptide region. Recent experiments using transient expression in HeLa cells (Weidemann, A., Konig, G., Bunke, D., Fischer, P., Salbaum, J.M., Masters, C.L., and Beyreuther, K. (1989) Cell 57, 115-126) indicate that the beta APP undergoes several posttranslational modifications including the cleavage and secretion of a large portion of its extracellular domain. The nature and fate of the fragment that remains cell-associated following this cleavage has not heretofore been described. The metabolism of this fragment may have particular significance in Alzheimer's disease since it must contain at least part of the beta-peptide. To study the metabolic fate of this fragment, we have established cell lines overexpressing the 695- and 751-amino acid versions of beta APP. Pulse-chase studies show that this system is similar to the HeLa cell system in that both proteins are synthesized first as membrane-bound proteins of approximately 98 and 108 kDa carrying asparagine-linked sugar side chains and are subsequently processed into higher molecular mass forms by the attachment of sulfate, phosphate, and further sugar groups including sialic acid, adding approximately 20 kDa in apparent molecular mass. The mature form of beta APP is cleaved and rapidly secreted, leaving an 11.5-kDa fragment with the transmembrane region and the cytoplasmic domain behind in the cell. This fragment is stable with a half-life of at least 4 h.
Oltersdorf
T
T
Athena Neurosciences Inc., South San Francisco, California 94080.
Ward
P J
PJ
Henriksson
T
T
Beattie
E C
EC
Neve
R
R
Lieberburg
I
I
Fritz
L C
LC
D000544:Alzheimer Disease / Q000378:metabolism*; D000682:Amyloid / Q000235:genetics / Q000378:metabolism*; D016564:Amyloid beta-Protein Precursor; D015153:Blotting, Western; D002461:Cell Line, Transformed; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D006207:Half-Life; D006801:Humans; D007150:Immunohistochemistry; D007163:Immunosorbent Techniques; D007700:Kinetics; D008970:Molecular Weight; D010446:Peptide Fragments / Q000378:metabolism*; D011498:Protein Precursors / Q000235:genetics / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D014162:Transfection; D014407:Tumor Cells, Cultured
D000682:Amyloid; D016564:Amyloid beta-Protein Precursor; D010446:Peptide Fragments; D011498:Protein Precursors; D004247:DNA