By Weiming Xia, Huaxi Xu

ISBN-10: 0849322456

ISBN-13: 9780849322457

Within the look for an efficient remedy for Alzheimer's disorder, APP is a special version protein that illustrates the big range of simple and complicated characterization innovations to be had.

Exploring quite a few organic options to explain the constitution and serve as of this transmembrane protein, this article provides each one strategy with unique, step by step protocols to accomplish reproducible effects and supply a framework for learning different membrane proteins.

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Extra resources for Amyloid precursor protein : a practical approach

Example text

The most highly conserved region of the molecule occurs at the N-terminal end, a cysteine-rich region that includes a heparin-binding domain (D1 or growth factor domain, GFD) and a metal-binding domain (D2 or copper binding domain, CuBD). All APP isoforms contain highly acidic domains; 45% of their residues are either Asp or Glu (D3). 1 Domain structure of APP. Domains are labeled D1 to D8 and the number of residues in each domain is indicated. The three-dimensional structure of GFD is shown as an alpha-carbon trace at the N-terminal end of the molecule.

The supernatant is loaded directly onto the heparin–hyperD column. 3. 0 M NaCl gradient in column equilibration buffer. 4. 12,13,25 5. 0. 6. 0. 7. The pooled fractions from the heparin column are loaded onto the QHyperD column and bound proteins eluted with a 50-ml linear gradient of NaCl (0 to 500 mM) in column equilibration buffer. 8. GFD elutes at a concentration of 50 mM NaCl. 9. 2). 10. The purified GFD is concentrated between 4 and 5 mg/ml for crystallization trials. Care must be taken to maintain the purified GFD at 4oC because the protein readily forms microcrystals if allowed to warm up.

C. Amyloid precursor protein modulates the interaction of nerve growth factor with p75 receptor and potentiates its activation of trkA phosphorylation. Mol. Brain. Res. 56, 125, 1998. 33. Henry A. et al. Inhibition of platelet activation by the Alzheimer’s disease amyloid precursor protein. Br. J. Haematol. 103, 402, 1998. 34. J. MOLSCRIPT: a program to produce both detailed and schematic plots of proteins. J. Appl. Crystallogr. 24, 946, 1991. fm Page 41 Wednesday, November 3, 2004 3:20 PM 4 Analysis of Amyloid Precursor Protein Processing Protease β-Secretase: Tools for Memapsin 2 (β-Secretase) Inhibition Studies Gerald Koelsch, Vajira Weerasena, Dongwoo Shin, Arun K.

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