Abstract
A pathological hallmark of many protein-misfolding diseases is the formation of insoluble aggregates. Quantitative methods are needed to better resolve and define the formation, aggregation, and temporal dynamics of soluble misfolded proteins in native settings. In this book chapter we describe simple and sensitive detection methods to characterize high ordered aggregates (AGERA) and subsets of distinct soluble aggregates (SEC-FRET) of mutant huntingtin protein in biological samples.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Chiti F, Dobson CM (2006) Protein misfolding, functional amyloid, and human disease. Annu Rev Biochem 75:333–366
Sarah L, Hands SL, Wyttenbach A (2010) Neurotoxic protein oligomerisation associated with polyglutamine diseases. Acta Neuropathol 120:419–437
Hazeki N, Tukamoto T, Gogo J et al (2000) Formic acid dissolves aggregates of an N-terminal huntingtin fragment containing an expanded polyglutamine tract: applying to quantification of protein components of the aggregates. Biochem Biophys Res Commun 277(2):386–393
Scherzinger E, Lurz R, Turmaine M et al (1997) Huntingtin-encoded polyglutamine expansions form amyloid-like protein aggregates in vitro and in vivo. Cell 90(3):549–558
Kryndushkin DS, Alexandrov IM, Ter-Avanesyan MD et al (2003) Yeast [PSI+] prion aggregates are formed by small Sup35 polymers fragmented by Hsp104. J Biol Chem 278(49):49636–49643
Bagriantsev SN, Kushnirov VV, Liebman SW (2006) Analysis of amyloid aggregates using agarose gel electrophoresis. Methods Enzymol 412:33–48
Ko J, Ou S, Patterson PH (2001) New anti-huntingtin monoclonal antibodies: implications for huntingtin conformation and its binding proteins. Brain Res Bull 56:319–329
Weiss A, Abramowski D, Bibel M et al (2009) Single-step detection of mutant Huntingtin in animal and human tissues: a bioassay for Huntington’s disease. Anal Biochem 395(1):8–15
Baldo B, Paganetti P, Grueninger S et al (2012) TR-FRET based duplex immunoassay reveals an inverse correlation of soluble and aggregated mutant huntingtin in Huntington’s disease. Chem Biol 19(2):264–275
Marcellin D, Abramowski D, Young D et al (2012) Fragments of HdhQ150 mutant huntingtin form a soluble oligomer pool that declines with aggregate deposition upon aging. PLoS One 7(9):e44457
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2013 Springer Science+Business Media, New York
About this protocol
Cite this protocol
Lotz, G.P., Weiss, A. (2013). Immuno-based Detection Assays to Quantify Distinct Mutant Huntingtin Conformations in Biological Samples. In: Hatters, D., Hannan, A. (eds) Tandem Repeats in Genes, Proteins, and Disease. Methods in Molecular Biology, vol 1017. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-438-8_12
Download citation
DOI: https://doi.org/10.1007/978-1-62703-438-8_12
Published:
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-437-1
Online ISBN: 978-1-62703-438-8
eBook Packages: Springer Protocols