Aggregation and Fibril Structure of AβM01-42 and Aβ1-42

Robert Silvers, Michael T. Colvin, Kendra K. Frederick, Angela C. Jacavone, Susan Lindquist, Sara Linse, Robert G. Griffin

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

A mechanistic understanding of Aβ aggregation and high-resolution structures of Aβ fibrils and oligomers are vital to elucidating relevant details of neurodegeneration in Alzheimer's disease, which will facilitate the rational design of diagnostic and therapeutic protocols. The most detailed and reproducible insights into structure and kinetics have been achieved using Aβ peptides produced by recombinant expression, which results in an additional methionine at the N-terminus. While the length of the C-terminus is well established to have a profound impact on the peptide's aggregation propensity, structure, and neurotoxicity, the impact of the N-terminal methionine on the aggregation pathways and structure is unclear. For this reason, we have developed a protocol to produce recombinant Aβ1-42, sans the N-terminal methionine, using an N-terminal small ubiquitin-like modifier-Aβ1-42 fusion protein in reasonable yield, with which we compared aggregation kinetics with AβM01-42 containing the additional methionine residue. The data revealed that Aβ1-42 and AβM01-42 aggregate with similar rates and by the same mechanism, in which the generation of new aggregates is dominated by secondary nucleation of monomers on the surface of fibrils. We also recorded magic angle spinning nuclear magnetic resonance spectra that demonstrated that excellent spectral resolution is maintained with both AβM01-42 and Aβ1-42 and that the chemical shifts are virtually identical in dipolar recoupling experiments that provide information about rigid residues. Collectively, these results indicate that the structure of the fibril core is unaffected by N-terminal methionine. This is consistent with the recent structures of AβM01-42 in which M0 is located at the terminus of a disordered 14-amino acid N-terminal tail.

Original languageEnglish (US)
Pages (from-to)4850-4859
Number of pages10
JournalBiochemistry
Volume56
Issue number36
DOIs
StatePublished - Sep 12 2017

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Methionine
Agglomeration
Peptides
Magic angle spinning
Kinetics
Spectral resolution
Chemical shift
Ubiquitin
Oligomers
Alzheimer Disease
Nucleation
Magnetic Resonance Spectroscopy
Fusion reactions
Monomers
Nuclear magnetic resonance
Amino Acids
Proteins
Experiments
Therapeutics

ASJC Scopus subject areas

  • Biochemistry

Cite this

Silvers, R., Colvin, M. T., Frederick, K. K., Jacavone, A. C., Lindquist, S., Linse, S., & Griffin, R. G. (2017). Aggregation and Fibril Structure of AβM01-42 and Aβ1-42. Biochemistry, 56(36), 4850-4859. https://doi.org/10.1021/acs.biochem.7b00729

Aggregation and Fibril Structure of AβM01-42 and Aβ1-42. / Silvers, Robert; Colvin, Michael T.; Frederick, Kendra K.; Jacavone, Angela C.; Lindquist, Susan; Linse, Sara; Griffin, Robert G.

In: Biochemistry, Vol. 56, No. 36, 12.09.2017, p. 4850-4859.

Research output: Contribution to journalArticle

Silvers, R, Colvin, MT, Frederick, KK, Jacavone, AC, Lindquist, S, Linse, S & Griffin, RG 2017, 'Aggregation and Fibril Structure of AβM01-42 and Aβ1-42', Biochemistry, vol. 56, no. 36, pp. 4850-4859. https://doi.org/10.1021/acs.biochem.7b00729
Silvers R, Colvin MT, Frederick KK, Jacavone AC, Lindquist S, Linse S et al. Aggregation and Fibril Structure of AβM01-42 and Aβ1-42. Biochemistry. 2017 Sep 12;56(36):4850-4859. https://doi.org/10.1021/acs.biochem.7b00729
Silvers, Robert ; Colvin, Michael T. ; Frederick, Kendra K. ; Jacavone, Angela C. ; Lindquist, Susan ; Linse, Sara ; Griffin, Robert G. / Aggregation and Fibril Structure of AβM01-42 and Aβ1-42. In: Biochemistry. 2017 ; Vol. 56, No. 36. pp. 4850-4859.
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