By Weiming Xia, Huaxi Xu
Within the look for an efficient therapy for Alzheimer's affliction, APP is a distinct version protein that illustrates the big choice of simple and complex characterization recommendations on hand.
Exploring various organic suggestions to explain the constitution and serve as of this transmembrane protein, this article offers every one process with designated, 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
The fourth and final is a morphological approach to examine APP internalization. 1,22 The 1G7 antibody is radioiodinated using IODO-GEN precoated iodination tubes (Pierce, Rockford, IL; #28601). fm Page 22 Wednesday, November 3, 2004 2:59 PM to the antibody. Excessive iodine is finally removed by gel filtration, typically with a disposable NAP-5 (Sephadex G-25) column (Amersham Biosciences, Piscataway, NJ; #17-0853-01). The entire procedure must be performed in an approved protective hood to avoid uptake of free radioactive iodine.
5). Sequence alignments of APP orthologues and paralogues show their GFD regions are well conserved with sequence identities ranging from 36 to 84%; all the cysteine residues are strictly conserved. This suggests that the fold described here is maintained across the APP family. 19 Maintenance of the disulfide bridge in this region is critical for neurite outgrowth19 and activation of MAP kinase,27 suggesting that the conformation of the loop is important. The surface is dominated by the β-hairpin loop (residues 96 to 110) representing the most mobile region of the structure.
Cell Sci. 109, 991, 1996. 20. Soriano, S. et al. Expression of beta-amyloid precursor protein-CD3gamma chimeras to demonstrate the selective generation of amyloid beta(1-40) and amyloid beta(142) peptides within secretory and endocytic compartments. J. Biol. Chem. 274, 32295, 1999. 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.