Abstract
The cleavage of amyloid precursor protein (APP) by γ-secretase produces Aβ peptides, which are prominent features in Alzheimer’s disease and have been extensively studied. By contrast, APP intracellular domain (AICD), also a product of this cleavage event, has received little or no investigative attention. A major reason for this is that AICD is generally not detected in tissue lysates and, therefore, is neglected as a non-relevant product of APP metabolism. However, recent studies have shown that AICD regulates a number of important cellular events. Furthermore, we found that contrary to previous assertions, AICD can be detected in brain lysates using Western blotting if an antigen retrieval protocol is employed. Here we describe the protocol for AICD detection and note the biological relevance of AICD in physiological and pathological conditions.
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References
Glenner, G. G., and Wong, C. W. (1984) Alzheimer’s disease and Down’s syndrome: sharing of a unique cerebrovascular amyloid fibril protein. Biochem Biophys Res Commun 122, 1131–5.
Hardy, J., and Selkoe, D. J. (2002) The amyloid hypothesis of Alzheimer’s disease: progress and problems on the road to therapeutics. Science 297, 353–6.
Hebert, S. S., Serneels, L., Tolia, A., Craessaerts, K., Derks, C., Filippov, M. A., Muller, U., and De Strooper, B. (2006) Regulated intramembrane proteolysis of amyloid precursor protein and regulation of expression of putative target genes. EMBO Rep 7, 739–45.
Muller, T., Meyer, H. E., Egensperger, R., and Marcus, K. (2008) The amyloid precursor protein intracellular domain (AICD) as modulator of gene expression, apoptosis, and cytoskeletal dynamics-relevance for Alzheimer’s disease. Prog Neurobiol 85, 393–406.
Ryan, K. A., and Pimplikar, S. W. (2005) J Cell Biol 171, 327–35.
Passer, B., Pellegrini, L., Russo, C., Siegel, R. M., Lenardo, M. J., Schettini, G., Bachmann, M., Tabaton, M., and D’Adamio, L. (2000) J Alzheimers Dis 2, 289–301.
Ma, Q. H., Futagawa, T., Yang, W. L., Jiang, X. D., Zeng, L., Takeda, Y., Xu, R. X., Bagnard, D., Schachner, M., Furley, A. J., Karagogeos, D., Watanabe, K., Dawe, G. S., and Xiao, Z. C. (2008) A TAG1-APP signalling pathway through Fe65 negatively modulates neurogenesis. Nat Cell Biol 10, 283–94.
Nakaya, T., and Suzuki, T. (2006) Role of APP phosphorylation in FE65-dependent gene transactivation mediated by AICD. Genes Cells 11, 633–45.
Pinnix, I., Musunuru, U., Tun, H., Sridharan, A., Golde, T., Eckman, C., Ziani-Cherif, C., Onstead, L., and Sambamurti, K. (2001) A novel gamma -secretase assay based on detection of the putative C-terminal fragment-gamma of amyloid beta protein precursor. J Biol Chem 276, 481–7.
Yu, C., Kim, S. H., Ikeuchi, T., Xu, H., Gasparini, L., Wang, R., and Sisodia, S. S. (2001) Characterization of a presenilin-mediated amyloid precursor protein carboxyl-terminal fragment gamma. Evidence for distinct mechanisms involved in gamma -secretase processing of the APP and Notch1 transmembrane domains. J Biol Chem 276, 43756–60.
Acknowledgments
The work in author’s laboratory is supported by grants from the NIH (R01-AG026146), Alzheimer’s Association, and Rotary-CART funds. We thank Dr. Chris Nelson for his comments on this manuscript.
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Pimplikar, S.W., Suryanarayana, A. (2010). Detection of APP Intracellular Domain in Brain Tissue. In: Roberson, E. (eds) Alzheimer's Disease and Frontotemporal Dementia. Methods in Molecular Biology, vol 670. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-744-0_7
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DOI: https://doi.org/10.1007/978-1-60761-744-0_7
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