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; Wennemers, Helma; Zenobi, Renato

doi:10.1039/c9sc03248g

Cited by 27 publications

(38 citation statements)

References 47 publications

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“…The duplex is denatured at 55 ± 1°C. Both thermal denaturation temperatures are very similar to those obtained by CD when monitoring specific wavelengths as previously demonstrated for other DNA complexes 10 and for collagen triple helices 63 (21°C at 269 nm and 53°C at 242 nm for the triplex-duplex and duplex-monomer transitions, respectively; Supplementary Fig. 12).…”

Section: Resultssupporting

confidence: 84%

Studying biomolecular folding and binding using temperature-jump mass spectrometry

et al. 2020

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Characterizing folding and complex formation of biomolecules provides a view into their thermodynamics, kinetics and folding pathways. Deciphering kinetic intermediates is particularly important because they can often be targeted by drugs. The key advantage of native mass spectrometry over conventional methods that monitor a single observable is its ability to identify and quantify coexisting species. Here, we show the design of a temperature-jump electrospray source for mass spectrometry that allows one to perform fast kinetics experiments (0.16-32 s) at different temperatures (10-90°C). The setup allows recording of both folding and unfolding kinetics by using temperature jumps from high to low, and low to high, temperatures. Six biological systems, ranging from peptides to proteins to DNA complexes, exemplify the use of this device. Using temperature-dependent experiments, the folding and unfolding of a DNA triplex are studied, providing detailed information on its thermodynamics and kinetics.

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Section: Resultssupporting

confidence: 84%

Studying biomolecular folding and binding using temperature-jump mass spectrometry

et al. 2020

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“…Protein melting (El‐Baba et al, 2018) or thermal heating of the solution (Wang, Bondarenko, & Kaltashov, 2018; Kohler et al, 2019) within the ESI emitter provide new approaches for determining the thermal stabilities of peptides, proteins, and even protein complexes. The combination of VT‐ESI with IM‐MS affords the ability to investigate structural transitions (refolded states) of biomolecules by kinetically trapping folding intermediates in the gas phase.…”

Section: Folding/refolding Proteins In Solution Using Vt‐esimentioning

confidence: 99%

“…The breadth of the studies described above clearly illustrate the unique potential for CIU, VT‐ESI‐IM‐MS, and MS/MS for studies of complex protein systems; however, incorporating additional MS‐based structural approaches adds to the biophysical characterization of proteins and protein complexes and their labeling with covalent and/or noncovalent modifiers. As examples, our current level of understanding of the structure(s) of partially metalated metallothionein comes from combining covalent labeling of the cysteine side chains with N ‐ethylmaleimide (Kohler et al, 2019) with top‐down and bottom‐up proteomics approaches (Chen, Russell, & Russell, 2013; Chen & Russell, 2015b).…”

Section: Combining Ciu and Vt‐esi‐im‐ms To Probe Stabilities Of Protementioning

confidence: 99%

The Ims Paradox: A Perspective on Structural Ion Mobility‐mass Spectrometry

McCabe

Hebert

Shirzadeh

et al. 2020

Mass Spectrometry Reviews

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Studies of large proteins, protein complexes, and membrane protein complexes pose new challenges, most notably the need for increased ion mobility (IM) and mass spectrometry (MS) resolution. This review covers evolutionary developments in IM‐MS in the authors' and key collaborators' laboratories with specific focus on developments that enhance the utility of IM‐MS for structural analysis. IM‐MS measurements are performed on gas phase ions, thus “structural IM‐MS” appears paradoxical—do gas phase ions retain their solution phase structure? There is growing evidence to support the notion that solution phase structure(s) can be retained by the gas phase ions. It should not go unnoticed that we use “structures” in this statement because an important feature of IM‐MS is the ability to deal with conformationally heterogeneous systems, thus providing a direct measure of conformational entropy. The extension of this work to large proteins and protein complexes has motivated our development of Fourier‐transform IM‐MS instruments, a strategy first described by Hill and coworkers in 1985 (Anal Chem, 1985, 57, pp. 402–406) that has proved to be a game‐changer in our quest to merge drift tube (DT) and ion mobility and the high mass resolution orbitrap MS instruments. DT‐IMS is the only method that allows first‐principles determinations of rotationally averaged collision cross sections (CSS), which is essential for studies of biomolecules where the conformational diversities of the molecule precludes the use of CCS calibration approaches. The Fourier transform‐IM‐orbitrap instrument described here also incorporates the full suite of native MS/IM‐MS capabilities that are currently employed in the most advanced native MS/IM‐MS instruments. © 2020 John Wiley & Sons Ltd. Mass Spec Rev

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“…[55] Contrary to CD,m ass spectrometry requires ionization and transfer of molecules into the gas phase where they are detected usually with an electron multiplier type detector.Our group recently demonstrated that native MS coupled with temperaturecontrolled nanoelectrospray (TCnESI) source is au seful analytical tools for structural biology and can retrieve detailed thermodynamic data on non-covalent complexes of peptides,p roteins,a nd oligonucleotides. [56,57] MS can be beneficial in studies focusing on multi-domain oligonucleotide complexes where the number of potential intermediates is unknown or the intermediates occur at low abundance.…”

Section: Introductionmentioning

confidence: 99%

Novel Insight into Proximal DNA Domain Interactions from Temperature‐Controlled Electrospray Ionization Mass Spectrometry

2021

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Quadruplexes are non-canonical nucleic acid structures essential for many cellular processes. Hybrid quadruplex-duplex oligonucleotide assemblies comprised of multiple domains are challenging to study with conventional biophysical methods due to their structural complexity.Here,weintroduce anovel method based on native mass spectrometry (MS) coupled with ac ustom-built temperature-controlled nanoelectrosprayi onization (TCnESI) source designed to investigate interactions between proximal DNAd omains.T hermal denaturation experiments were aimed to study unfolding of multi-stranded oligonucleotide constructs derived from biologically relevant structures and to identify unfolding intermediates.Using the TCnESI MS,weobserved changes in T m and thermodynamic characteristics of proximal DNAdomains depending on the number of domains, their position, and order in asingle experiment.

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

Abstract: Temperature-programmed native electrospray ionization mass spectrometry gives detailed insight into the assembly of model collagen triple helices.

Cited by 27 publications

References 47 publications

Studying biomolecular folding and binding using temperature-jump mass spectrometry

Studying biomolecular folding and binding using temperature-jump mass spectrometry

The Ims Paradox: A Perspective on Structural Ion Mobility‐mass Spectrometry

Novel Insight into Proximal DNA Domain Interactions from Temperature‐Controlled Electrospray Ionization Mass Spectrometry

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