Protein kinase C regulation of intracellular and cell surface amyloid precursor protein (APP) cleavage in CHO695 cells.
Cleavage of amyloid precursor protein (APP) by beta-secretase generates beta-amyloid (Abeta), the major component of senile plaques in Alzheimer's disease. Cleavage of APP by alpha-secretase prevents Abeta formation, producing nonamyloidogenic secreted APPs products. PKC-regulated APP alpha-secretase cleavage has been shown to involve tumor necrosis factor alpha (TNF-alpha) converting enzyme (TACE). To determine the location of APP cleavage, we examined PKC-regulated APPs secretion by examining cell surface versus intracellular APP in CHO cells stably expressing APP(695) (CHO695). We demonstrate that PKC regulates cell surface and intracellular APP cleavage. The majority of secreted APPs originates from the intracellular compartment, and PKC does not cause an increase in APP trafficking to the cell surface for cleavage. Therefore, intracellular APP regulated by PKC must be cleaved at an intracellular site. Experiments utilizing Brefeldin A suggest APP cleavage occurs at the Golgi or late in the secretory pathway. Experiments using TAPI, an inhibitor of TACE, demonstrate PKC-regulated APPs secretion from the cell surface is inhibited after pretreatment with TAPI, and APPs secretion from the intracellular pool is partially inhibited after pretreatment with TAPI. These findings suggest PKC-regulated APP cleavage occurs at multiple locations within the cell and both events appear to involve TACE.
Jolly-Tornetta
C
C
Department of Pathology and Laboratory Medicine, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
Wolf
B A
BA
D051722:ADAM Proteins; D000072198:ADAM17 Protein; D016564:Amyloid beta-Protein Precursor / Q000378:metabolism*; D000818:Animals; D020126:Brefeldin A / Q000494:pharmacology; D016466:CHO Cells; D002451:Cell Compartmentation; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D004151:Dipeptides / Q000494:pharmacology; D006056:Golgi Apparatus / Q000378:metabolism; D006877:Hydroxamic Acids / Q000494:pharmacology; D008565:Membrane Proteins; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors; D011493:Protein Kinase C / Q000378:metabolism*; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D013755:Tetradecanoylphorbol Acetate
D016564:Amyloid beta-Protein Precursor; D004151:Dipeptides; D006877:Hydroxamic Acids; D008565:Membrane Proteins; C092152:N-((2-(hydroxyaminocarbonyl)methyl)-4-methylpentanoyl)-3-(2'-naphthyl)alanylalanine, 2-aminoethylamide; D020126:Brefeldin A; D011493:Protein Kinase C; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000072198:ADAM17 Protein; D013755:Tetradecanoylphorbol Acetate