Abstract
SummaryQuantitative mass spectrometry has established proteome-wide regulation of protein abundance and post-translational modifications in various biological processes. Here, we used quantitative mass spectrometry to systematically analyze the thermal stability and solubility of proteins on a proteome-wide scale during the eukaryotic cell cycle. We demonstrate pervasive variation of these biophysical parameters with most changes occurring in mitosis and G1. Various cellular pathways and components vary in thermal stability, such as cell-cycle factors, polymerases, and chromatin remodelers. We demonstrate that protein thermal stability serves as a proxy for enzyme activity, DNA binding, and complex formation in situ. Strikingly, a large cohort of intrinsically disordered and mitotically phosphorylated proteins is stabilized and solubilized in mitosis, suggesting a fundamental remodeling of the biophysical environment of the mitotic cell. Our data represent a rich resource for cell, structural, and systems biologists interested in proteome regulation during biological transitions.
Highlights
Mass spectrometry (MS)-based proteomics is a powerful method to accurately quantify distinct features of proteomes (Mann et al, 2013) and has provided comprehensive insight into the variability of protein abundance under distinct biological conditions, including different cell types (Geiger et al, 2012), protein complexes (Havugimana et al, 2012), and subcellular fractions (Boisvert et al, 2010)
Intact cells were heated and lysed with a mild detergent (NP-40), and soluble proteins were identified by multiplexed quantitative MS based on tandem mass tag (TMT) labeling (Werner et al, 2014)
RNA Polymerase II Stability Correlates with Transcriptional Activity we investigated whether our data capture changes in protein-chromatin interactions
Summary
Mass spectrometry (MS)-based proteomics is a powerful method to accurately quantify distinct features of proteomes (Mann et al, 2013) and has provided comprehensive insight into the variability of protein abundance under distinct biological conditions, including different cell types (Geiger et al, 2012), protein complexes (Havugimana et al, 2012), and subcellular fractions (Boisvert et al, 2010). Recent advances in MS multiplexing technologies (Werner et al, 2014) enable the global measurement of protein melting curves (thermal proteome profiling; TPP) (Savitski et al, 2014). It is well established that various aspects of proteome organization, including protein abundance and post-translational modifications, vary during cellcycle progression (Dephoure et al, 2008; Olsen et al, 2010). We hypothesized that cell-cycle-dependent post-translational modifications, protein-protein interactions, and spatial rearrangements to distinct biophysical environments globally influence protein thermal stability (Jensen et al, 2006; Jongsma et al, 2015; Olsen et al, 2010; Pelisch et al, 2014). We systematically measured in situ protein thermal stability, abundance, and solubility during cell-cycle progression on a proteome-wide scale
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
