Receptor- and non-receptor tyrosine kinases induce processing of the amyloid precursor protein: role of the low-density lipoprotein receptor-related protein.
The Alzheimer's beta-amyloid peptides derive from the proteolytic processing of the beta-amyloid precursor protein, APP, by beta- and gamma-secretases. The regulation of this processing is not fully understood. Experimental evidence suggests that the activation of pathways involving protein tyrosine kinases, such as PDGFR and Src, could induce the cleavage of APP and in turn the generation of amyloid peptides. In this paper we addressed the effect of receptor and nonreceptor protein tyrosine kinases on the cleavage of APP and the mechanisms of their action. To this aim, we developed an in vitro system based on the APP-Gal4 fusion protein stably transfected in SHSY5Y neuroblastoma cell line. The cleavage of this molecule, induced by various stimuli, results in the activation of the transcription of the luciferase gene under the control of Gal4 cis-elements. By using this experimental system we demonstrated that, similarly to Src, three tyrosine kinases, TrkA, Ret and EGFR, induced the cleavage of APP-Gal4. We excluded that this effect was mediated by the activation of Ras-MAPK, PI3K-Akt and PLC-gamma pathways. Furthermore, the direct phosphorylation of the APP cytosolic domain does not affect Abeta peptide generation. On the contrary, experiments in cells lacking the LDL-receptor related protein LRP support the hypothesis that the interaction of APP with LRP is required for the induction of APP cleavage by tyrosine kinases.
Minopoli
Giuseppina
G
CEINGE Biotecnologie Avanzate, Dipartimento di Biochimica e Biotecnologie Mediche, Università di Napoli Federico II, Napoli, Italia.
Passaro
Fabiana
F
Aloia
Luigi
L
Carlomagno
Francesca
F
Melillo
Rosa Marina
RM
Santoro
Massimo
M
Forzati
Floriana
F
Zambrano
Nicola
N
Russo
Tommaso
T
D016564:Amyloid beta-Protein Precursor / Q000187:drug effects* / Q000378:metabolism*; D002461:Cell Line, Transformed; D045744:Cell Line, Tumor; D005998:Glycine / Q000378:metabolism; D006801:Humans; D047468:Immunoprecipitation; D020794:Receptor Protein-Tyrosine Kinases / Q000494:pharmacology*; D011973:Receptors, LDL / Q000502:physiology*; D014162:Transfection / Q000379:methods
D016564:Amyloid beta-Protein Precursor; D011973:Receptors, LDL; D020794:Receptor Protein-Tyrosine Kinases; D005998:Glycine