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Detection and Quantification of Protein-Protein Interactions by Mass Spectrometry
- Abstract:
Protein interaction networks supply the fundamental means for biological pathways to complete tasks, allowing cells to perform basic functions that sustain life and promote growth. This work describes the detection and quantification of Saccharomyces cerevisiae HSP90 protein-protein interactions using SPROX (Stability of Proteins from Rates of Oxidation), a mass spectrometry-based chemical modification strategy that probes protein stability. In two SPROX experiments, six proteins were identified as potential HSP90 binding partners, four of which have been previously documented to interact with HSP90 in the literature. These six identified proteins bound HSP90 with Kd values ranging from 300 nM to 1.6 μM. The application of SPROX for protein-protein interactions provides a complementary method that can be used in concert with conventional techniques to detect and quantify protein-protein interactions.
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