An intein-based strategy for the production of tag-free huntingtin exon 1 proteins enables new insights into the polyglutamine dependence of Httex1 aggregation and fibril formation

Sophie Vieweg, Annalisa Ansaloni, Zhe Ming Wang, John B. Warner, Hilal A. Lashuel

Research output: Contribution to journalArticle

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Abstract

The first exon of the Huntingtin protein (Httex1) is one of the most actively studied Htt fragments because its overexpression in R6/2 transgenic mice has been shown to recapitulate several key features of Huntington disease. However, the majority of biophysical studies of Httex1 are based on assessing the structure and aggregation of fusion constructs where Httex1 is fused to large proteins, such as glutathione S-transferase, maltosebinding protein, or thioredoxin, or released in solution upon in situ cleavage of these proteins. Herein, we report an inteinbased strategy that allows, for the first time, the rapid and efficient production of native tag-free Httex1 with polyQ repeats ranging from 7Q to 49Q. Aggregation studies on these proteins enabled us to identify interesting polyQ-length-dependent effects on Httex1 oligomer and fibril formation that were previously not observed using Httex1 fusion proteins or Httex1 proteins produced by in situ cleavage of fusion proteins. Our studies revealed the inability of Httex1-7Q/15Q to undergo amyloid fibril formation and an inverse correlation between fibril length and polyQ repeat length, suggesting possible polyQ length-dependent differences in the structural properties of the Httex1 aggregates. Altogether, our findings underscore the importance of working with tag-free Httex1 proteins and indicate that model systems based on non-native Httex1 sequences may not accurately reproduce the effect of polyQ repeat length and solution conditions on Httex1 aggregation kinetics and structural properties.

Original languageEnglish
Pages (from-to)12074-12086
Number of pages13
JournalJournal of Biological Chemistry
Volume291
Issue number23
DOIs
Publication statusPublished - 2016

Fingerprint

Inteins
Exons
Agglomeration
Proteins
Fusion reactions
Structural properties
Thioredoxins
polyglutamine
Huntington Disease
Glutathione Transferase
Amyloid
Transgenic Mice
Oligomers

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

An intein-based strategy for the production of tag-free huntingtin exon 1 proteins enables new insights into the polyglutamine dependence of Httex1 aggregation and fibril formation. / Vieweg, Sophie; Ansaloni, Annalisa; Wang, Zhe Ming; Warner, John B.; Lashuel, Hilal A.

In: Journal of Biological Chemistry, Vol. 291, No. 23, 2016, p. 12074-12086.

Research output: Contribution to journalArticle

Vieweg, Sophie ; Ansaloni, Annalisa ; Wang, Zhe Ming ; Warner, John B. ; Lashuel, Hilal A. / An intein-based strategy for the production of tag-free huntingtin exon 1 proteins enables new insights into the polyglutamine dependence of Httex1 aggregation and fibril formation. In: Journal of Biological Chemistry. 2016 ; Vol. 291, No. 23. pp. 12074-12086.
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