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Purification and Aggregation of the Amyloid Precursor Protein Intracellular Domain
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Purification and Aggregation of the Amyloid Precursor Protein Intracellular Domain

Purification and Aggregation of the Amyloid Precursor Protein Intracellular Domain

DOI:
10.3791/4204-v

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10:08 min

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August 28, 2012

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, , , ,
1Department of Surgery,University of Texas Medical Branch , 2Department of Neuroscience and Cell Biology,University of Texas Medical Branch

Chapters

  • 00:05Title
  • 01:35Expression of Recombinant APP Intracellular Domain
  • 02:55Purification of GST-AICD
  • 05:23Thrombin Cleavage of GST-AICD and Purification of AICD
  • 06:54AICD Aggregation for Atomic Force Microscopy
  • 07:56Representative AICD Aggregation Results
  • 09:32Conclusion

Summary

Automatic Translation

Automatic Translation

A method for large-scale purification of the APP intracellular domain (AICD) is described. We also describe methodology to induce in vitro AICD aggregation and visualization by atomic force microscopy. The methods described are useful for biochemical/structural characterization of the AICD and the effects of molecular chaperones on its aggregation.

Tags

Amyloid Precursor ProteinAPPAlzheimer’s DiseaseAICDProteasesα-secretaseβ-secretaseγ-secretaseAβ PeptideSenile PlaquesFe65Tip60Transcription RegulationProtein-protein InteractionsHolo-APPC-terminal FragmentsAggregationMolecular Chaperone Ubiquilin-1Hydrophobic DomainsIntrinsically DisorderedStable Secondary Structure

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