ac1c00772_si_003.xlsx (131.24 kB)
Download fileThermal Analysis of a Mixture of Ribosomal Proteins by vT-ESI-MS: Toward a Parallel Approach for Characterizing the Stabilitome
dataset
posted on 2021-06-08, 19:38 authored by Tarick
J. El-Baba, Shannon A. Raab, Rachel P. Buckley, Christopher J. Brown, Corinne A. Lutomski, Lucas W. Henderson, Daniel W. Woodall, Jiangchuan Shen, Jonathan C. Trinidad, Hengyao Niu, Martin F. Jarrold, David H. Russell, Arthur Laganowsky, David E. ClemmerThe thermal stabilities of endogenous,
intact proteins and protein
assemblies in complex mixtures were characterized in parallel by means
of variable-temperature electrospray ionization coupled to mass spectrometry
(vT-ESI-MS). The method is demonstrated by directly measuring the
melting transitions of seven proteins from a mixture of proteins derived
from ribosomes. A proof-of-concept measurement of a fraction of an Escherichia coli lysate is provided to extend this
approach to characterize the thermal stability of a proteome. As the
solution temperature is increased, proteins and protein complexes
undergo structural and organizational transitions; for each species,
the folded ↔ unfolded and assembled ↔ disassembled populations
are monitored based on changes in vT-ESI-MS charge state distributions
and masses. The robustness of the approach illustrates a step toward
the proteome-wide characterization of thermal stabilities and structural
transitionsthe stabilitome.