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

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Abstract

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.

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

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Publication details

The article was received on 02 Jul 2019, accepted on 29 Aug 2019 and first published on 03 Sep 2019


Article type: Edge Article
DOI: 10.1039/C9SC03248G
Chem. Sci., 2019, Advance Article
  • Open access: Creative Commons BY-NC license
    All publication charges for this article have been paid for by the Royal Society of Chemistry

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

    M. Köhler, A. Marchand, N. B. Hentzen, J. Egli, A. I. Begley, H. Wennemers and R. Zenobi, Chem. Sci., 2019, Advance Article , DOI: 10.1039/C9SC03248G

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