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Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers

Collagen model peptides are useful for understanding the assembly and structure of collagen triple helices. The design of self-assembling heterotrimeric helices is particularly challenging and often affords mixtures of non-covalent assemblies that are difficult to characterize by conventional NMR an...

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Autores principales: Köhler, Martin, Marchand, Adrien, Hentzen, Nina B., Egli, Jasmine, Begley, Alina I., Wennemers, Helma, Zenobi, Renato
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Royal Society of Chemistry 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6977553/
https://www.ncbi.nlm.nih.gov/pubmed/32015805
http://dx.doi.org/10.1039/c9sc03248g
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author Köhler, Martin
Marchand, Adrien
Hentzen, Nina B.
Egli, Jasmine
Begley, Alina I.
Wennemers, Helma
Zenobi, Renato
author_facet Köhler, Martin
Marchand, Adrien
Hentzen, Nina B.
Egli, Jasmine
Begley, Alina I.
Wennemers, Helma
Zenobi, Renato
author_sort Köhler, Martin
collection PubMed
description Collagen model peptides are useful for understanding the assembly and structure of collagen triple helices. The design of self-assembling heterotrimeric helices is particularly challenging and often affords mixtures of non-covalent assemblies that are difficult to characterize by conventional NMR and CD spectroscopic techniques. This can render a detailed understanding of the factors that control heterotrimer formation difficult and restrict rational design. Here, we present a novel method based on electrospray ionization mass spectrometry to investigate homo- and heterotrimeric collagen model peptides. Under native conditions, the high resolving power of mass spectrometry was used to access the stoichiometric composition of different triple helices in complex mixtures. A temperature-controlled electrospray ionization source was built to perform thermal denaturation experiments and provided melting temperatures of triple helices. These were found to be in good agreement with values obtained from CD spectroscopic measurements. Importantly, for mixtures of coexisting homo- and heterotrimers, which are difficult to analyze by conventional methods, our technique allowed for the identification and monitoring of the unfolding of each individual species. Their respective melting temperatures could easily be accessed in a single experiment, using small amounts of sample.
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spelling pubmed-69775532020-02-03 Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers Köhler, Martin Marchand, Adrien Hentzen, Nina B. Egli, Jasmine Begley, Alina I. Wennemers, Helma Zenobi, Renato Chem Sci Chemistry Collagen model peptides are useful for understanding the assembly and structure of collagen triple helices. The design of self-assembling heterotrimeric helices is particularly challenging and often affords mixtures of non-covalent assemblies that are difficult to characterize by conventional NMR and CD spectroscopic techniques. This can render a detailed understanding of the factors that control heterotrimer formation difficult and restrict rational design. Here, we present a novel method based on electrospray ionization mass spectrometry to investigate homo- and heterotrimeric collagen model peptides. Under native conditions, the high resolving power of mass spectrometry was used to access the stoichiometric composition of different triple helices in complex mixtures. A temperature-controlled electrospray ionization source was built to perform thermal denaturation experiments and provided melting temperatures of triple helices. These were found to be in good agreement with values obtained from CD spectroscopic measurements. Importantly, for mixtures of coexisting homo- and heterotrimers, which are difficult to analyze by conventional methods, our technique allowed for the identification and monitoring of the unfolding of each individual species. Their respective melting temperatures could easily be accessed in a single experiment, using small amounts of sample. Royal Society of Chemistry 2019-09-03 /pmc/articles/PMC6977553/ /pubmed/32015805 http://dx.doi.org/10.1039/c9sc03248g Text en This journal is © The Royal Society of Chemistry 2019 http://creativecommons.org/licenses/by-nc/3.0/ This article is freely available. This article is licensed under a Creative Commons Attribution Non Commercial 3.0 Unported Licence (CC BY-NC 3.0)
spellingShingle Chemistry
Köhler, Martin
Marchand, Adrien
Hentzen, Nina B.
Egli, Jasmine
Begley, Alina I.
Wennemers, Helma
Zenobi, Renato
Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
title Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
title_full Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
title_fullStr Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
title_full_unstemmed Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
title_short Temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
title_sort temperature-controlled electrospray ionization mass spectrometry as a tool to study collagen homo- and heterotrimers
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6977553/
https://www.ncbi.nlm.nih.gov/pubmed/32015805
http://dx.doi.org/10.1039/c9sc03248g
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