Nanodisc Reconstitution and Characterization of Amyloid-β Precursor Protein C99

Area of Science:

• Biochemistry and structural biology
• Neuroscience and neurodegenerative disease research

Background:

• Alzheimer's disease (AD) pathology is closely linked to the amyloid precursor protein (APP).
• Amyloid-β peptide production from APP fragmentation initiates amyloid toxicity in AD.
• Understanding APP structure and dynamics in lipid bilayers is critical but challenging.

Purpose of the Study:

• To reconstitute the C-terminal domain of APP into a stable, membrane-mimicking environment.
• To characterize the structure and dynamics of reconstituted APP using advanced biophysical techniques.
• To demonstrate the feasibility of using polymer nanodiscs for APP investigation.

Main Methods:

• Reconstitution of the 99-residue C-terminal domain of APP into polymer nanodiscs.
• Characterization using size-exclusion chromatography and mass spectrometry.
• Structural and dynamic analysis via solution NMR and magic-angle spinning solid-state NMR.

Main Results:

• Successful reconstitution of the APP C-terminal domain into polymer nanodiscs.
• Comprehensive biophysical characterization confirming successful integration and stability.
• Demonstration of polymer nanodiscs as a viable tool for studying APP in membrane environments.

Conclusions:

• Polymer nanodiscs provide a suitable platform for studying APP structure and dynamics.
• This method facilitates investigation of APP in a near-native lipid-bilayer environment.
• The approach is extendable to characterizing APP within native Escherichia coli membranes.