Synapse-binding subpopulations of Aβ oligomers sensitive to peptide assembly blockers and scFv antibodies

Pauline T. Velasco, Marie Heffern, Adriano Sebollela, Izolda A. Popova, Pascale N. Lacor, Kevin B. Lee, Xiaoxia Sun, Benjamin N. Tiano, Kirsten L. Viola, Amanda L. Eckermann, Thomas J. Meade, William L. Klein

Research output: Contribution to journalArticle

22 Citations (Scopus)

Abstract

Amyloid β42 self-assembly is complex, with multiple pathways leading to large insoluble fibrils or soluble oligomers. Oligomers are now regarded as most germane to Alzheimers pathogenesis. We have investigated the hypothesis that oligomer formation itself occurs through alternative pathways, with some leading to synapse-binding toxins. Immediately after adding synthetic peptide to buffer, solutions of Aβ42 were separated by a 50 kDa filter and fractions assessed by SDS-PAGE silver stain, Western blot, immunoprecipitation, and capacity for synaptic binding. Aβ42 rapidly assembled into aqueous-stable oligomers, with similar protein abundance in small (<50 kDa) and large (>50 kDa) oligomer fractions. Initially, both fractions were SDS-labile and resolved into tetramers, trimers, and monomers by SDS-PAGE. Upon continued incubation, the larger oligomers developed a small population of SDS-stable 10-16mers, and the smaller oligomers generated gel-impermeant complexes. The two fractions associated differently with neurons, with prominent synaptic binding limited to larger oligomers. Even within the family of larger oligomers, synaptic binding was associated with only a subset of these species, as a new scFv antibody (NUsc1) immunoprecipitated only a small portion of the oligomers while eliminating synaptic binding. Interestingly, low doses of the peptide KLVFFA blocked assembly of the 10-16mers, and this result was associated with loss of the smaller clusters of oligomers observed at synaptic sites. What distinguishes these smaller clusters from the unaffected larger clusters is not yet known. Results indicate that distinct species of Aβ oligomers are generated by alternative assembly pathways and that synapse-binding subpopulations of Aβ oligomers could be specifically targeted for Alzheimers therapeutics.

Original languageEnglish (US)
Pages (from-to)972-981
Number of pages10
JournalACS Chemical Neuroscience
Volume3
Issue number11
DOIs
StatePublished - Nov 21 2012
Externally publishedYes

Fingerprint

Single-Chain Antibodies
Oligomers
Synapses
Polyacrylamide Gel Electrophoresis
Peptides
Population Dynamics
Silver
Immunoprecipitation
Amyloid
Buffers
Coloring Agents
Western Blotting
Gels
Neurons
Proteins
Therapeutics
Self assembly

Keywords

  • Alzheimers disease
  • amyloid beta
  • blocking peptide
  • oligomerization
  • single-chain variable fragment antibody
  • therapeutics

ASJC Scopus subject areas

  • Biochemistry
  • Physiology
  • Cognitive Neuroscience
  • Cell Biology

Cite this

Synapse-binding subpopulations of Aβ oligomers sensitive to peptide assembly blockers and scFv antibodies. / Velasco, Pauline T.; Heffern, Marie; Sebollela, Adriano; Popova, Izolda A.; Lacor, Pascale N.; Lee, Kevin B.; Sun, Xiaoxia; Tiano, Benjamin N.; Viola, Kirsten L.; Eckermann, Amanda L.; Meade, Thomas J.; Klein, William L.

In: ACS Chemical Neuroscience, Vol. 3, No. 11, 21.11.2012, p. 972-981.

Research output: Contribution to journalArticle

Velasco, PT, Heffern, M, Sebollela, A, Popova, IA, Lacor, PN, Lee, KB, Sun, X, Tiano, BN, Viola, KL, Eckermann, AL, Meade, TJ & Klein, WL 2012, 'Synapse-binding subpopulations of Aβ oligomers sensitive to peptide assembly blockers and scFv antibodies', ACS Chemical Neuroscience, vol. 3, no. 11, pp. 972-981. https://doi.org/10.1021/cn300122k
Velasco, Pauline T. ; Heffern, Marie ; Sebollela, Adriano ; Popova, Izolda A. ; Lacor, Pascale N. ; Lee, Kevin B. ; Sun, Xiaoxia ; Tiano, Benjamin N. ; Viola, Kirsten L. ; Eckermann, Amanda L. ; Meade, Thomas J. ; Klein, William L. / Synapse-binding subpopulations of Aβ oligomers sensitive to peptide assembly blockers and scFv antibodies. In: ACS Chemical Neuroscience. 2012 ; Vol. 3, No. 11. pp. 972-981.
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