Jump to main content
Jump to site search
Access to RSC content Close the message box

Continue to access RSC content when you are not at your institution. Follow our step-by-step guide.


Issue 20, 2015
Previous Article Next Article

Comparing equilibrium and kinetic protein unfolding using time-resolved electrospray-coupled ion mobility mass spectrometry

Author affiliations

Abstract

Protein unfolding intermediates are thought to play a critical role in conformational pathogenesis, acting as a ‘gateway’ to inactivation or pathogenic aggregation. Unfolding intermediates have long been studied either by populating partially-folded species at equilibrium using incresingly denaturing conditions, or by transiently populating ‘kinetic’ intermediates under fully denaturing conditions using a time-resolved approach (e.g. stopped-flow fluorescence). However, it is not clear that the folding intermediates populated under equilibrium conditions are comparable to intermediates transiently populated in kinetic experiments. In this work, we combine time-resolved electrospray (TRESI) with travelling wave Ion Mobility Spectrometry (IMS) for the first time to directly compare equilibrium and kinetic unfolding intermediates of cytochrome c. Our results show a high degree of correlation between all species populated under these substantially different regimes.

Graphical abstract: Comparing equilibrium and kinetic protein unfolding using time-resolved electrospray-coupled ion mobility mass spectrometry

Back to tab navigation

Article information


Submitted
28 Apr 2015
Accepted
22 Jun 2015
First published
22 Jun 2015

Analyst, 2015,140, 6973-6979
Article type
Paper
Author version available

Comparing equilibrium and kinetic protein unfolding using time-resolved electrospray-coupled ion mobility mass spectrometry

P. Liuni, B. Deng and D. J. Wilson, Analyst, 2015, 140, 6973
DOI: 10.1039/C5AN00843C

Social activity

Search articles by author

Spotlight

Advertisements