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Updated: Jun 5, 2026
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Generation of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Using a SUMO Fusion Strategy
Published on: June 27, 2018
Assessing mutant huntingtin fragment and polyglutamine aggregation by atomic force microscopy.
Kathleen A Burke1, Jordan Godbey, Justin Legleiter
1The C. Eugene Bennett Department of Chemistry, West Virginia University, 217 Clark Hall, P.O. Box 6045, Morgantown, WV 26506, USA.
Methods (San Diego, Calif.)
|December 29, 2010
Summary
Huntington disease research uses atomic force microscopy (AFM) to study mutant huntingtin (htt) protein aggregation. This method visualizes fibrillar aggregates, aiding in understanding disease mechanisms and identifying distinct aggregate species.
Area of Science:
- Neuroscience
- Biochemistry
- Biophysics
Background:
- Huntington disease (HD) is a neurodegenerative disorder.
- HD is caused by expanded polyglutamine (polyQ) repeats in the huntingtin (htt) protein.
- Expanded polyQ domains lead to htt aggregation and inclusion body formation, characteristic of HD.
Purpose of the Study:
- To present protocols for studying mutant htt fragment and polyQ peptide aggregation using atomic force microscopy (AFM).
- To enable characterization of aggregate formation over time and distinguish specific aggregate species.
- To facilitate dynamic studies of aggregation processes and aggregate interactions.
Main Methods:
- Ex situ AFM to analyze aggregate formation during protein incubation.
- Morphological feature analysis for distinguishing aggregate species in heterogeneous reactions.
- Time-lapse AFM in solution for real-time tracking of synthetic polyQ peptide aggregation.
Main Results:
- AFM protocols effectively characterize htt aggregate formation and morphology.
- Methods allow for unambiguous identification of specific aggregate species.
- Time-lapse AFM provides dynamic insights into polyQ peptide aggregation and interactions.
Conclusions:
- AFM is a powerful tool for studying the aggregation kinetics and morphology of htt fragments and polyQ peptides relevant to Huntington disease.
- The presented methods enhance the ability to analyze complex aggregation reactions and observe dynamic processes.
- This research contributes to a deeper understanding of the molecular mechanisms underlying Huntington disease pathogenesis.
