Preferential adsorption, internalization and resistance to degradation of the major isoform of the Alzheimer's amyloid peptide, A beta 1-42, in differentiated PC12 cells.
A central question in Alzheimer's disease (AD) is the role of amyloid in pathogenesis. Recent discoveries implicating the longer A beta 1-42 form of amyloid in pathogenesis led us to characterize the interaction of A beta with cells to elucidate differences that might account for these observations. We characterized the adsorption, internalization and degradation of radiolabeled A beta in NGF-differentiated PC12 cells under conditions that are not acutely toxic. All A beta peptides examined absorb to the surface of PC12 cells and are internalized; however the adsorption and internalization of A beta 1-42 is significantly greater than that of A beta 1-40 and A beta 1-28. The adsorption of A beta 1-42 is decreased by treatment of the cells with neuraminidase, but not heparitinase. The fate of the internalized A beta 1-42 is also very different than shorter A beta peptides; a fraction of the internalized A beta 1-42 accumulates intracellularly and is resistant to degradation for at least 3 days while A beta 1-40 and shorter peptides are eliminated with a half life of about 1 h. A beta 1-42 does not appear to inhibit lysosomal hydrolases, since A beta 1-28 is degraded at the same rate in the presence or absence of A beta 1-42. The intracellular A beta 1-42 is located in a dense organellar compartment and colocalizes with the lysosomal markers Lucifer Yellow and horseradish peroxidase. These data indicate that there are significant differences in the cell surface adsorption, internalization and catabolism of A beta 1-42 compared to A beta 1-40 and A beta 1-28. These differences may be important for the preferential accumulation of the longer A beta 1-42 isoform and its association with AD pathogenesis.
Burdick
D
D
Department of Molecular Biology, University of California, Irvine 92697, USA.
Kosmoski
J
J
Knauer
M F
MF
Glabe
C G
CG
D000327:Adsorption; D000595:Amino Acid Sequence; D016564:Amyloid beta-Protein Precursor / Q000737:chemistry / Q000378:metabolism* / Q000493:pharmacokinetics; D000818:Animals; D002451:Cell Compartmentation / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D011992:Endosomes / Q000737:chemistry; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D007536:Isomerism; D007546:Isoquinolines; D008247:Lysosomes / Q000737:chemistry; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D009410:Nerve Degeneration / Q000502:physiology; D016716:PC12 Cells / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats
D016564:Amyloid beta-Protein Precursor; D005456:Fluorescent Dyes; D007546:Isoquinolines; C017475:lucifer yellow