Initial protein unfolding events revealed by 213 nm UVPD coupled to IM-MS

06 April 2018, Version 1
This content is a preprint and has not undergone peer review at the time of posting.

Abstract

In this work we couple UVPD with activated ion mobility mass spectrometry to measure how three model proteins start to unfold. Ubiquitin, cytochrome c and myoglobin ions produced via nESI from salty solutions are subjected to UV irradiation pre-mobility separation, experiments are conducted with a range of source conditions which alter the conformation of the precursor ion as shown by the drift time profiles. For all three proteins the compact structures result in less fragmentation than more extended structures which emerge following progressive in-source activation. Cleavage sites are found to differ between conformational ensembles, for example, for the dominant charge state of cytochrome c [M+7H]7+, cleavage at Phe10, Thr19 and Val20 was only observed in activating conditions while cleavage at Ala43 is dramatically enhanced. Mapping the photo-cleaved fragments onto crystallographic structures provides insight into the local structural changes that occur as protein unfolding progresses, which is coupled to global restructuring observed in the drift time profiles.

Keywords

IM-MS
Ion mobility
Mass spectrometry
Native mass spectrometry
213 nm
UVPD
Photodissociation
Protein unfolding
activated IM-MS
Collision-Induced Unfolding
nano electrospray
Cytochrome c
Ubiquitin
Myoglobin

Supplementary materials

Title
Description
Actions
Title
Theisen 213nm UVPD SI
Description
Actions

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