Conformational Dynamics of Biomolecules by Trapped Ion Mobility Spectrometry Dynamics

Molano-Arévalo, Juan Camilo

doi:10.25148/etd.fidc006557

Cited by 2 publications

(1 citation statement)

References 3 publications

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“…However, for the 9 + charge state, all solution conditions analyzed under harsh parameters exhibit much more fragmentation than any of the solution states fragmented by ECD with gentle instrument conditions. This is likely due to the elimination of intramolecular interactions due to extensive ion heating [50], which was also observed for the ATDs under these harsh conditions. Cytochrome c 9 + ECD sequence coverage increases from 54% in water to 72% at pH 2.35 and pH 2.19 as the ion heating further unfolds the protein compared to only solution unfolding before analysis with ion transmission under gentle conditions (10%, 15%, and 35% respectively).…”

Section: Ecd Under Gentle Conditions Figure 4: Ecd Sequence Coverage ...mentioning

confidence: 64%

Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions

Cain

Webb

2023

Anal Bioanal Chem

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Electrospray ionization mass spectrometry (ESI-MS) experiments, including ion mobilitymass spectrometry (ESI-IM-MS), and electron capture dissociation (ECD) of proteins ionized from aqueous solutions, have been used for the study of solution-like structures of intact proteins. By mixing aqueous proteins with denaturants online before ESI, the amount of protein unfolding can be precisely controlled and rapidly analyzed, permitting the characterization of protein folding intermediates in protein folding pathways. Herein, we mixed various pH solutions online with aqueous cytochrome C for unfolding and characterizing its unfolding intermediates with ESI-MS charge state distribution measurements, ion mobility, and ECD. The presence of folding intermediates and unfolded cytochrome c structures were detected from changes in charge states, arrival time distributions (ATDs), and ECD fragmentation. We also compared structures from nondenaturing and denaturing solution mixtures measured under "gentle" (i.e., low energy) ion transmission conditions with structures measured under "harsh" (higher energy) transmission. This work confirms that when using "gentle" instrument conditions, the gas-phase cytochrome c ions reflect attributes of the various solution-phase structures. However, "harsh" conditions that maximize ion transmission produce annealed, extended structures that no longer correlate with changes in solution structure.

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Section: Ecd Under Gentle Conditions Figure 4: Ecd Sequence Coverage ...mentioning

confidence: 64%

Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions

Cain

Webb

2023

Anal Bioanal Chem

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Online Protein Unfolding Characterized by Ion Mobility Electron Capture Dissociation Mass Spectrometry: Cytochrome C from Neutral and Acidic Solutions

Cain

Webb

2022

Preprint

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Electrospray ionization mass spectrometry (ESI-MS) experiments, including ion mobility-mass spectrometry (ESI-IM-MS), and electron capture dissociation (ECD) of proteins ionized from aqueous solutions, have been used for the study of solution-like structures of intact proteins. By mixing aqueous proteins with denaturants online before ESI, the amount of protein unfolding can be precisely controlled and rapidly analyzed, permitting the characterization of protein folding intermediates in protein folding pathways. Herein, we mixed various pH solutions online with aqueous cytochrome C for unfolding and characterizing its unfolding intermediates with ESI-MS charge state distribution measurements, ion mobility, and ECD. The presence of folding intermediates and unfolded cytochrome c structures were detected from changes in charge states, arrival time distributions (ATDs), and ECD fragmentation. We also compared structures from nondenaturing and denaturing solution mixtures measured under “gentle” (i.e., low energy) ion transmission conditions with structures measured under “harsh” (higher energy) transmission. This work confirms that when using “gentle” instrument conditions, the gas-phase cytochrome c ions reflect attributes of the various solution-phase structures. However, “harsh” conditions that maximize ion transmission produce annealed, extended structures that no longer correlate with changes in solution structure.

show abstract

Conformational Dynamics of Biomolecules by Trapped Ion Mobility Spectrometry Dynamics

Cited by 2 publications

References 3 publications

Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions

Online protein unfolding characterized by ion mobility electron capture dissociation mass spectrometry: cytochrome C from neutral and acidic solutions

Online Protein Unfolding Characterized by Ion Mobility Electron Capture Dissociation Mass Spectrometry: Cytochrome C from Neutral and Acidic Solutions

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