By Weiming Xia, Huaxi Xu
Within the look for a good remedy for Alzheimer's disorder, APP is a different version protein that illustrates the wide variety of simple and complex characterization suggestions to be had.
Exploring various organic thoughts to elucidate the constitution and serve as of this transmembrane protein, this article offers every one strategy with exact, step by step protocols to accomplish reproducible effects and supply a framework for learning different membrane proteins.
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Additional info for Amyloid precursor protein : a practical approach
21. U. et al. The cytoplasmic domain of the LDL receptor-related protein regulates multiple steps in APP processing. EMBO J. 21, 5691, 2002. 22. H. Trafficking of cell surface beta-amyloid precursor protein: retrograde and transcytotic transport in cultured neurons. J. Cell Biol. 129, 431, 1995. 23. A. A. Rab15 mediates an early endocytic event in Chinese hamster ovary cells. J. Biol. Chem. 274, 22303, 1999. fm Page 29 Wednesday, November 3, 2004 2:59 PM 3 Strategies for Crystallizing the N-Terminal Growth Factor Domain of Amyloid Precursor Protein William J.
3 KINETICS OF SECRETION AND ENDOCYTOSIS OF APP This protocol examines the trafficking of APP by pulse/chase analysis. A population of surface APP molecules is initially bound to radiolabeled antibody at 4˚C and is then, after removal of unbound antibody, allowed to transit when rewarmed to 37˚C. At various time points, the supernatant containing the secreted APP is collected and any remaining antibody is removed from the cell surface by acid washes. Finally, the cell pellet is lysed (acid resistant fraction) to obtain the internalized APP fraction and all samples are measured in a gamma counter.
Crystal structure of the N-terminal, growth factor-like domain of Alzheimer amyloid precursor protein. Nature Struct. Biol. 6, 327, 1999. 25. Hilbich, C. et al. Amyloid-like properties of peptides flanking the epitope of amyloid precursor protein-specific monoclonal antibody 22C11. J. Biol. Chem. 268, 26571, 1993. 26. , Crystallization of Biological Macromolecules, Cold Spring Harbor Laboratory Press, New York, 1999. 27. M. et al. Amino-terminal region of the beta-amyloid precursor protein activates mitogen-activated protein kinase.
Amyloid precursor protein : a practical approach by Weiming Xia, Huaxi Xu