Cargando…

Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation

[Image: see text] Small organic molecules that inhibit functional bacterial amyloid fibers, curli, are promising new antibiotics. Here we investigated the mechanism by which the ring-fused 2-pyridone FN075 inhibits fibrillation of the curli protein CsgA. Using a variety of biophysical techniques, we...

Descripción completa

Detalles Bibliográficos
Autores principales: Horvath, Istvan, Weise, Christoph F., Andersson, Emma K., Chorell, Erik, Sellstedt, Magnus, Bengtsson, Christoffer, Olofsson, Anders, Hultgren, Scott J., Chapman, Matthew, Wolf-Watz, Magnus, Almqvist, Fredrik, Wittung-Stafshede, Pernilla
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2012
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3290101/
https://www.ncbi.nlm.nih.gov/pubmed/22260746
http://dx.doi.org/10.1021/ja209829m
_version_ 1782224945220157440
author Horvath, Istvan
Weise, Christoph F.
Andersson, Emma K.
Chorell, Erik
Sellstedt, Magnus
Bengtsson, Christoffer
Olofsson, Anders
Hultgren, Scott J.
Chapman, Matthew
Wolf-Watz, Magnus
Almqvist, Fredrik
Wittung-Stafshede, Pernilla
author_facet Horvath, Istvan
Weise, Christoph F.
Andersson, Emma K.
Chorell, Erik
Sellstedt, Magnus
Bengtsson, Christoffer
Olofsson, Anders
Hultgren, Scott J.
Chapman, Matthew
Wolf-Watz, Magnus
Almqvist, Fredrik
Wittung-Stafshede, Pernilla
author_sort Horvath, Istvan
collection PubMed
description [Image: see text] Small organic molecules that inhibit functional bacterial amyloid fibers, curli, are promising new antibiotics. Here we investigated the mechanism by which the ring-fused 2-pyridone FN075 inhibits fibrillation of the curli protein CsgA. Using a variety of biophysical techniques, we found that FN075 promotes CsgA to form off-pathway, non-amyloidogenic oligomeric species. In light of the generic properties of amyloids, we tested whether FN075 would also affect the fibrillation reaction of human α-synuclein, an amyloid-forming protein involved in Parkinson’s disease. Surprisingly, FN075 stimulates α-synuclein amyloid fiber formation as measured by thioflavin T emission, electron microscopy (EM), and atomic force microscopy (AFM). NMR data on (15)N-labeled α-synuclein show that upon FN075 addition, α-synuclein oligomers with 7 nm radius form in which the C-terminal 40 residues remain disordered and solvent exposed. The polypeptides in these oligomers contain β-like secondary structure, and the oligomers are detectable by AFM, EM, and size-exclusion chromatography (SEC). Taken together, FN075 triggers oligomer formation of both proteins: in the case of CsgA, the oligomers do not proceed to fibers, whereas for α-synuclein, the oligomers are poised to rapidly form fibers. We conclude that there is a fine balance between small-molecule inhibition and templation that depends on protein chemistry.
format Online
Article
Text
id pubmed-3290101
institution National Center for Biotechnology Information
language English
publishDate 2012
publisher American Chemical Society
record_format MEDLINE/PubMed
spelling pubmed-32901012012-02-29 Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation Horvath, Istvan Weise, Christoph F. Andersson, Emma K. Chorell, Erik Sellstedt, Magnus Bengtsson, Christoffer Olofsson, Anders Hultgren, Scott J. Chapman, Matthew Wolf-Watz, Magnus Almqvist, Fredrik Wittung-Stafshede, Pernilla J Am Chem Soc [Image: see text] Small organic molecules that inhibit functional bacterial amyloid fibers, curli, are promising new antibiotics. Here we investigated the mechanism by which the ring-fused 2-pyridone FN075 inhibits fibrillation of the curli protein CsgA. Using a variety of biophysical techniques, we found that FN075 promotes CsgA to form off-pathway, non-amyloidogenic oligomeric species. In light of the generic properties of amyloids, we tested whether FN075 would also affect the fibrillation reaction of human α-synuclein, an amyloid-forming protein involved in Parkinson’s disease. Surprisingly, FN075 stimulates α-synuclein amyloid fiber formation as measured by thioflavin T emission, electron microscopy (EM), and atomic force microscopy (AFM). NMR data on (15)N-labeled α-synuclein show that upon FN075 addition, α-synuclein oligomers with 7 nm radius form in which the C-terminal 40 residues remain disordered and solvent exposed. The polypeptides in these oligomers contain β-like secondary structure, and the oligomers are detectable by AFM, EM, and size-exclusion chromatography (SEC). Taken together, FN075 triggers oligomer formation of both proteins: in the case of CsgA, the oligomers do not proceed to fibers, whereas for α-synuclein, the oligomers are poised to rapidly form fibers. We conclude that there is a fine balance between small-molecule inhibition and templation that depends on protein chemistry. American Chemical Society 2012-01-19 2012-02-22 /pmc/articles/PMC3290101/ /pubmed/22260746 http://dx.doi.org/10.1021/ja209829m Text en Copyright © 2012 American Chemical Society http://pubs.acs.org This is an open-access article distributed under the ACS AuthorChoice Terms & Conditions. Any use of this article, must conform to the terms of that license which are available at http://pubs.acs.org.
spellingShingle Horvath, Istvan
Weise, Christoph F.
Andersson, Emma K.
Chorell, Erik
Sellstedt, Magnus
Bengtsson, Christoffer
Olofsson, Anders
Hultgren, Scott J.
Chapman, Matthew
Wolf-Watz, Magnus
Almqvist, Fredrik
Wittung-Stafshede, Pernilla
Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
title Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
title_full Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
title_fullStr Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
title_full_unstemmed Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
title_short Mechanisms of Protein Oligomerization: Inhibitor of Functional Amyloids Templates α-Synuclein Fibrillation
title_sort mechanisms of protein oligomerization: inhibitor of functional amyloids templates α-synuclein fibrillation
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3290101/
https://www.ncbi.nlm.nih.gov/pubmed/22260746
http://dx.doi.org/10.1021/ja209829m
work_keys_str_mv AT horvathistvan mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT weisechristophf mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT anderssonemmak mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT chorellerik mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT sellstedtmagnus mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT bengtssonchristoffer mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT olofssonanders mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT hultgrenscottj mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT chapmanmatthew mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT wolfwatzmagnus mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT almqvistfredrik mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation
AT wittungstafshedepernilla mechanismsofproteinoligomerizationinhibitoroffunctionalamyloidstemplatesasynucleinfibrillation