A novel crosslinking protocol stabilizes amyloid β oligomers capable of inducing Alzheimer's-associated pathologies

Erika N. Cline, Arighno Das, Maíra Assunção Bicca, Saad N. Mohammad, Luis F. Schachner, Josette M. Kamel, Nadia DiNunno, Anthea Weng, Jacob D. Paschall, Riana Lo Bu, Faraz M. Khan, Madeline G. Rollins, Ashley N. Ives, Gajendra S Shekhawat, Nilson Nunes-Tavares, Fernando G. de Mello, Philip D Compton, Neil L Kelleher, William L Klein*

*Corresponding author for this work

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Amyloid β oligomers (AβOs) accumulate early in Alzheimer's disease (AD) and experimentally cause memory dysfunction and the major pathologies associated with AD, for example, tau abnormalities, synapse loss, oxidative damage, and cognitive dysfunction. In order to develop the most effective AβO-targeting diagnostics and therapeutics, the AβO structures contributing to AD-associated toxicity must be elucidated. Here, we investigate the structural properties and pathogenic relevance of AβOs stabilized by the bifunctional crosslinker 1,5-difluoro-2,4-dinitrobenzene (DFDNB). We find that DFDNB stabilizes synthetic Aβ in a soluble oligomeric conformation. With DFDNB, solutions of Aβ that would otherwise convert to large aggregates instead yield solutions of stable AβOs, predominantly in the 50–300 kDa range, that are maintained for at least 12 days at 37°C. Structures were determined by biochemical and native top–down mass spectrometry analyses. Assayed in neuronal cultures and i.c.v.-injected mice, the DFDNB-stabilized AβOs were found to induce tau hyperphosphorylation, inhibit choline acetyltransferase, and provoke neuroinflammation. Most interestingly, DFDNB crosslinking was found to stabilize an AβO conformation particularly potent in inducing memory dysfunction in mice. Taken together, these data support the utility of DFDNB crosslinking as a tool for stabilizing pathogenic AβOs in structure-function studies. (Figure presented.).

Original languageEnglish (US)
Pages (from-to)822-836
Number of pages15
JournalJournal of neurochemistry
Volume148
Issue number6
DOIs
StatePublished - Mar 1 2019

Fingerprint

Pathology
Oligomers
Amyloid
Crosslinking
Alzheimer Disease
Conformations
Data storage equipment
Choline O-Acetyltransferase
Synapses
Mass spectrometry
Toxicity
Structural properties
Mass Spectrometry
2,4-dinitrobenzene

Keywords

  • Alzheimer's disease
  • DFDNB
  • amyloid beta oligomers
  • crosslinking
  • structure–function relationships

ASJC Scopus subject areas

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Cline, Erika N. ; Das, Arighno ; Bicca, Maíra Assunção ; Mohammad, Saad N. ; Schachner, Luis F. ; Kamel, Josette M. ; DiNunno, Nadia ; Weng, Anthea ; Paschall, Jacob D. ; Bu, Riana Lo ; Khan, Faraz M. ; Rollins, Madeline G. ; Ives, Ashley N. ; Shekhawat, Gajendra S ; Nunes-Tavares, Nilson ; de Mello, Fernando G. ; Compton, Philip D ; Kelleher, Neil L ; Klein, William L. / A novel crosslinking protocol stabilizes amyloid β oligomers capable of inducing Alzheimer's-associated pathologies. In: Journal of neurochemistry. 2019 ; Vol. 148, No. 6. pp. 822-836.
@article{e7f4bdf0792648a3b97d63e3050adb18,
title = "A novel crosslinking protocol stabilizes amyloid β oligomers capable of inducing Alzheimer's-associated pathologies",
abstract = "Amyloid β oligomers (AβOs) accumulate early in Alzheimer's disease (AD) and experimentally cause memory dysfunction and the major pathologies associated with AD, for example, tau abnormalities, synapse loss, oxidative damage, and cognitive dysfunction. In order to develop the most effective AβO-targeting diagnostics and therapeutics, the AβO structures contributing to AD-associated toxicity must be elucidated. Here, we investigate the structural properties and pathogenic relevance of AβOs stabilized by the bifunctional crosslinker 1,5-difluoro-2,4-dinitrobenzene (DFDNB). We find that DFDNB stabilizes synthetic Aβ in a soluble oligomeric conformation. With DFDNB, solutions of Aβ that would otherwise convert to large aggregates instead yield solutions of stable AβOs, predominantly in the 50–300 kDa range, that are maintained for at least 12 days at 37°C. Structures were determined by biochemical and native top–down mass spectrometry analyses. Assayed in neuronal cultures and i.c.v.-injected mice, the DFDNB-stabilized AβOs were found to induce tau hyperphosphorylation, inhibit choline acetyltransferase, and provoke neuroinflammation. Most interestingly, DFDNB crosslinking was found to stabilize an AβO conformation particularly potent in inducing memory dysfunction in mice. Taken together, these data support the utility of DFDNB crosslinking as a tool for stabilizing pathogenic AβOs in structure-function studies. (Figure presented.).",
keywords = "Alzheimer's disease, DFDNB, amyloid beta oligomers, crosslinking, structure–function relationships",
author = "Cline, {Erika N.} and Arighno Das and Bicca, {Ma{\'i}ra Assun{\cc}{\~a}o} and Mohammad, {Saad N.} and Schachner, {Luis F.} and Kamel, {Josette M.} and Nadia DiNunno and Anthea Weng and Paschall, {Jacob D.} and Bu, {Riana Lo} and Khan, {Faraz M.} and Rollins, {Madeline G.} and Ives, {Ashley N.} and Shekhawat, {Gajendra S} and Nilson Nunes-Tavares and {de Mello}, {Fernando G.} and Compton, {Philip D} and Kelleher, {Neil L} and Klein, {William L}",
year = "2019",
month = "3",
day = "1",
doi = "10.1111/jnc.14647",
language = "English (US)",
volume = "148",
pages = "822--836",
journal = "Journal of Neurochemistry",
issn = "0022-3042",
publisher = "Wiley-Blackwell",
number = "6",

}

Cline, EN, Das, A, Bicca, MA, Mohammad, SN, Schachner, LF, Kamel, JM, DiNunno, N, Weng, A, Paschall, JD, Bu, RL, Khan, FM, Rollins, MG, Ives, AN, Shekhawat, GS, Nunes-Tavares, N, de Mello, FG, Compton, PD, Kelleher, NL & Klein, WL 2019, 'A novel crosslinking protocol stabilizes amyloid β oligomers capable of inducing Alzheimer's-associated pathologies', Journal of neurochemistry, vol. 148, no. 6, pp. 822-836. https://doi.org/10.1111/jnc.14647

A novel crosslinking protocol stabilizes amyloid β oligomers capable of inducing Alzheimer's-associated pathologies. / Cline, Erika N.; Das, Arighno; Bicca, Maíra Assunção; Mohammad, Saad N.; Schachner, Luis F.; Kamel, Josette M.; DiNunno, Nadia; Weng, Anthea; Paschall, Jacob D.; Bu, Riana Lo; Khan, Faraz M.; Rollins, Madeline G.; Ives, Ashley N.; Shekhawat, Gajendra S; Nunes-Tavares, Nilson; de Mello, Fernando G.; Compton, Philip D; Kelleher, Neil L; Klein, William L.

In: Journal of neurochemistry, Vol. 148, No. 6, 01.03.2019, p. 822-836.

Research output: Contribution to journalArticle

TY - JOUR

T1 - A novel crosslinking protocol stabilizes amyloid β oligomers capable of inducing Alzheimer's-associated pathologies

AU - Cline, Erika N.

AU - Das, Arighno

AU - Bicca, Maíra Assunção

AU - Mohammad, Saad N.

AU - Schachner, Luis F.

AU - Kamel, Josette M.

AU - DiNunno, Nadia

AU - Weng, Anthea

AU - Paschall, Jacob D.

AU - Bu, Riana Lo

AU - Khan, Faraz M.

AU - Rollins, Madeline G.

AU - Ives, Ashley N.

AU - Shekhawat, Gajendra S

AU - Nunes-Tavares, Nilson

AU - de Mello, Fernando G.

AU - Compton, Philip D

AU - Kelleher, Neil L

AU - Klein, William L

PY - 2019/3/1

Y1 - 2019/3/1

N2 - Amyloid β oligomers (AβOs) accumulate early in Alzheimer's disease (AD) and experimentally cause memory dysfunction and the major pathologies associated with AD, for example, tau abnormalities, synapse loss, oxidative damage, and cognitive dysfunction. In order to develop the most effective AβO-targeting diagnostics and therapeutics, the AβO structures contributing to AD-associated toxicity must be elucidated. Here, we investigate the structural properties and pathogenic relevance of AβOs stabilized by the bifunctional crosslinker 1,5-difluoro-2,4-dinitrobenzene (DFDNB). We find that DFDNB stabilizes synthetic Aβ in a soluble oligomeric conformation. With DFDNB, solutions of Aβ that would otherwise convert to large aggregates instead yield solutions of stable AβOs, predominantly in the 50–300 kDa range, that are maintained for at least 12 days at 37°C. Structures were determined by biochemical and native top–down mass spectrometry analyses. Assayed in neuronal cultures and i.c.v.-injected mice, the DFDNB-stabilized AβOs were found to induce tau hyperphosphorylation, inhibit choline acetyltransferase, and provoke neuroinflammation. Most interestingly, DFDNB crosslinking was found to stabilize an AβO conformation particularly potent in inducing memory dysfunction in mice. Taken together, these data support the utility of DFDNB crosslinking as a tool for stabilizing pathogenic AβOs in structure-function studies. (Figure presented.).

AB - Amyloid β oligomers (AβOs) accumulate early in Alzheimer's disease (AD) and experimentally cause memory dysfunction and the major pathologies associated with AD, for example, tau abnormalities, synapse loss, oxidative damage, and cognitive dysfunction. In order to develop the most effective AβO-targeting diagnostics and therapeutics, the AβO structures contributing to AD-associated toxicity must be elucidated. Here, we investigate the structural properties and pathogenic relevance of AβOs stabilized by the bifunctional crosslinker 1,5-difluoro-2,4-dinitrobenzene (DFDNB). We find that DFDNB stabilizes synthetic Aβ in a soluble oligomeric conformation. With DFDNB, solutions of Aβ that would otherwise convert to large aggregates instead yield solutions of stable AβOs, predominantly in the 50–300 kDa range, that are maintained for at least 12 days at 37°C. Structures were determined by biochemical and native top–down mass spectrometry analyses. Assayed in neuronal cultures and i.c.v.-injected mice, the DFDNB-stabilized AβOs were found to induce tau hyperphosphorylation, inhibit choline acetyltransferase, and provoke neuroinflammation. Most interestingly, DFDNB crosslinking was found to stabilize an AβO conformation particularly potent in inducing memory dysfunction in mice. Taken together, these data support the utility of DFDNB crosslinking as a tool for stabilizing pathogenic AβOs in structure-function studies. (Figure presented.).

KW - Alzheimer's disease

KW - DFDNB

KW - amyloid beta oligomers

KW - crosslinking

KW - structure–function relationships

UR - http://www.scopus.com/inward/record.url?scp=85060749937&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85060749937&partnerID=8YFLogxK

U2 - 10.1111/jnc.14647

DO - 10.1111/jnc.14647

M3 - Article

VL - 148

SP - 822

EP - 836

JO - Journal of Neurochemistry

JF - Journal of Neurochemistry

SN - 0022-3042

IS - 6

ER -