Proteome Integral Solubility Alteration: A High-Throughput Proteomics Assay for Target Deconvolution

Various agents, including
drugs as well as nonmolecular stimuli,
induce alterations in the physicochemical properties of proteins in
cell lysates, living cells, and organisms. These alterations can be
probed by applying a stability- and solubility-modifying factor, such
as elevated temperature, to a varying degree. As a second dimension
of variation, drug concentration or agent intensity/concentration
can be used. Compared to standard approaches where curves are fitted
to protein solubility data acquired at different temperatures and
drug concentrations, Proteome Integral Solubility Alteration (PISA)
assay increases the analysis throughput by 1 to 2 orders of magnitude
for an unlimited number of factor variation points in such a scheme.
The consumption of the compound and biological material decreases
in PISA by the same factor. We envision widespread use of the PISA
approach in chemical biology and drug development.