Cargando…

Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation

The human molecular chaperone protein DNAJB6 was recently found to inhibit the formation of amyloid fibrils from polyglutamine peptides associated with neurodegenerative disorders such as Huntington disease. We show in the present study that DNAJB6 also inhibits amyloid formation by an even more agg...

Descripción completa

Detalles Bibliográficos
Autores principales: Månsson, Cecilia, Arosio, Paolo, Hussein, Rasha, Kampinga, Harm H., Hashem, Reem M., Boelens, Wilbert C., Dobson, Christopher M., Knowles, Tuomas P. J., Linse, Sara, Emanuelsson, Cecilia
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4223311/
https://www.ncbi.nlm.nih.gov/pubmed/25217638
http://dx.doi.org/10.1074/jbc.M114.595124
_version_ 1782343180717391872
author Månsson, Cecilia
Arosio, Paolo
Hussein, Rasha
Kampinga, Harm H.
Hashem, Reem M.
Boelens, Wilbert C.
Dobson, Christopher M.
Knowles, Tuomas P. J.
Linse, Sara
Emanuelsson, Cecilia
author_facet Månsson, Cecilia
Arosio, Paolo
Hussein, Rasha
Kampinga, Harm H.
Hashem, Reem M.
Boelens, Wilbert C.
Dobson, Christopher M.
Knowles, Tuomas P. J.
Linse, Sara
Emanuelsson, Cecilia
author_sort Månsson, Cecilia
collection PubMed
description The human molecular chaperone protein DNAJB6 was recently found to inhibit the formation of amyloid fibrils from polyglutamine peptides associated with neurodegenerative disorders such as Huntington disease. We show in the present study that DNAJB6 also inhibits amyloid formation by an even more aggregation-prone peptide (the amyloid-beta peptide, Aβ42, implicated in Alzheimer disease) in a highly efficient manner. By monitoring fibril formation using Thioflavin T fluorescence and far-UV CD spectroscopy, we have found that the aggregation of Aβ42 is retarded by DNAJB6 in a concentration-dependent manner, extending to very low sub-stoichiometric molar ratios of chaperone to peptide. Quantitative kinetic analysis and immunochemistry studies suggest that the high inhibitory efficiency is due to the interactions of the chaperone with aggregated forms of Aβ42 rather than the monomeric form of the peptide. This interaction prevents the growth of such species to longer fibrils and inhibits the formation of new amyloid fibrils through both primary and secondary nucleation. A low dissociation rate of DNAJB6 from Aβ42 aggregates leads to its incorporation into growing fibrils and hence to its gradual depletion from solution with time. When DNAJB6 is eventually depleted, fibril proliferation takes place, but the inhibitory activity can be prolonged by introducing DNAJB6 at regular intervals during the aggregation reaction. These results reveal the highly efficacious mode of action of this molecular chaperone against protein aggregation, and demonstrate that the role of molecular chaperones can involve interactions with multiple aggregated species leading to the inhibition of both principal nucleation pathways through which aggregates are able to form.
format Online
Article
Text
id pubmed-4223311
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher American Society for Biochemistry and Molecular Biology
record_format MEDLINE/PubMed
spelling pubmed-42233112014-11-07 Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation Månsson, Cecilia Arosio, Paolo Hussein, Rasha Kampinga, Harm H. Hashem, Reem M. Boelens, Wilbert C. Dobson, Christopher M. Knowles, Tuomas P. J. Linse, Sara Emanuelsson, Cecilia J Biol Chem Molecular Biophysics The human molecular chaperone protein DNAJB6 was recently found to inhibit the formation of amyloid fibrils from polyglutamine peptides associated with neurodegenerative disorders such as Huntington disease. We show in the present study that DNAJB6 also inhibits amyloid formation by an even more aggregation-prone peptide (the amyloid-beta peptide, Aβ42, implicated in Alzheimer disease) in a highly efficient manner. By monitoring fibril formation using Thioflavin T fluorescence and far-UV CD spectroscopy, we have found that the aggregation of Aβ42 is retarded by DNAJB6 in a concentration-dependent manner, extending to very low sub-stoichiometric molar ratios of chaperone to peptide. Quantitative kinetic analysis and immunochemistry studies suggest that the high inhibitory efficiency is due to the interactions of the chaperone with aggregated forms of Aβ42 rather than the monomeric form of the peptide. This interaction prevents the growth of such species to longer fibrils and inhibits the formation of new amyloid fibrils through both primary and secondary nucleation. A low dissociation rate of DNAJB6 from Aβ42 aggregates leads to its incorporation into growing fibrils and hence to its gradual depletion from solution with time. When DNAJB6 is eventually depleted, fibril proliferation takes place, but the inhibitory activity can be prolonged by introducing DNAJB6 at regular intervals during the aggregation reaction. These results reveal the highly efficacious mode of action of this molecular chaperone against protein aggregation, and demonstrate that the role of molecular chaperones can involve interactions with multiple aggregated species leading to the inhibition of both principal nucleation pathways through which aggregates are able to form. American Society for Biochemistry and Molecular Biology 2014-11-07 2014-09-12 /pmc/articles/PMC4223311/ /pubmed/25217638 http://dx.doi.org/10.1074/jbc.M114.595124 Text en © 2014 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version full access. Creative Commons Attribution Unported License (http://creativecommons.org/licenses/by/3.0/) applies to Author Choice Articles
spellingShingle Molecular Biophysics
Månsson, Cecilia
Arosio, Paolo
Hussein, Rasha
Kampinga, Harm H.
Hashem, Reem M.
Boelens, Wilbert C.
Dobson, Christopher M.
Knowles, Tuomas P. J.
Linse, Sara
Emanuelsson, Cecilia
Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation
title Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation
title_full Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation
title_fullStr Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation
title_full_unstemmed Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation
title_short Interaction of the Molecular Chaperone DNAJB6 with Growing Amyloid-beta 42 (Aβ42) Aggregates Leads to Sub-stoichiometric Inhibition of Amyloid Formation
title_sort interaction of the molecular chaperone dnajb6 with growing amyloid-beta 42 (aβ42) aggregates leads to sub-stoichiometric inhibition of amyloid formation
topic Molecular Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4223311/
https://www.ncbi.nlm.nih.gov/pubmed/25217638
http://dx.doi.org/10.1074/jbc.M114.595124
work_keys_str_mv AT manssoncecilia interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT arosiopaolo interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT husseinrasha interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT kampingaharmh interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT hashemreemm interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT boelenswilbertc interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT dobsonchristopherm interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT knowlestuomaspj interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT linsesara interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation
AT emanuelssoncecilia interactionofthemolecularchaperonednajb6withgrowingamyloidbeta42ab42aggregatesleadstosubstoichiometricinhibitionofamyloidformation