High throughput experimental determination of mini-protein local stability and fluctuation

Proteins are highly dynamic molecules characterized not only by global unfolding events but by localized fluctuations. These local fluctuations can drastically influence the binding affinity, overall stability, and function of proteins. While many biophysical methods exist to study protein stability, techniques that can provide residue-resolution measurements in high throughput are limited. The Rocklin lab has recently developed a high throughput hydrogen deuterium exchange mass spectrometry method that utilizes three dimensions of separation, liquid chromatography, ion mobility, and mass spectrometry, to assay the overall energy landscapes of thousands of intact mini-proteins in parallel.