Fig. 5.

The TPCA workflow. TPCA can be performed on intact cells or cell lysate. Lysed samples are first divided into an equal amount of aliquots and subjected to heat treatment with an increasing temperature gradient. Heat treatment induces denaturation and coaggregation of interacting proteins, which then co-precipitate. Upon centrifugation, the supernatant consisting of soluble proteins from different temperature treatment is retrieved for isobaric TMT-labelling and quantitative LC–MS/MS analysis. The abundance of each soluble proteins identified and quantified is then plotted against the temperatures to generate the “protein melting curve”