Publication
Title
Fixed-charge trimethyl pyrilium modification for enabling enhanced top-down mass spectrometry sequencing of intact protein complexes
Author
Abstract
Mass spectrometry of intact proteins and protein complexes has the potential to provide a trans formative level of information on biological systems, ranging from sequence and post-translational modification analysis to the structures of whole protein assemblies. This ambitious goal requires the efficient fragmentation of both intact proteins and the macromolecular, multicomponent machines they collaborate to create through noncovalent interactions. Improving technologies in an effort to achieve such fragmentation remains perhaps the greatest challenge facing current efforts to comprehensively analyze cellular protein composition and is essential to realizing the full potential of proteomics. In this work, we describe the use of a trimethyl pyrylium (TMP) fixed-charge covalent labeling strategy aimed at enhancing fragmentation challenging proteins and intact protein complexes. Combining analysis of TMP-modified and unmodified protein complexes results in a greater diversity of regions within the protein that give rise to fragments, and results in an up to 2.5-fold increase in sequence coverage when compared to unmodified protein alone, for protein complexes up to 148 kDa. TMP modification offers a simple and powerful platform to expand the capabilities of existing mass spectrometric instrumentation for the complete characterization of intact protein assemblies.
Language
English
Source (journal)
Analytical chemistry. - Washington, D.C., 1948, currens
Publication
Washington, D.C. : 2018
ISSN
0003-2700 [print]
5206-882X [online]
Volume/pages
90 :4 (2018) , p. 2756-2764
ISI
000426143100051
Pubmed ID
29360341
Full text (Publisher's DOI)
Full text (open access)
UAntwerpen
Faculty/Department
Research group
Publication type
Subject
Affiliation
Publications with a UAntwerp address
External links
Web of Science
Record
Identification
Creation 29.03.2018
Last edited 20.09.2021
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