Pinpointing changes in higher-order protein structure by hydrogen/deuterium exchange coupled to electron transfer dissociation mass spectrometry

This Feature describes the use of electron transfer dissociation (ETD) to analyze the hydrogen/deuterium exchange (HDX) of proteins at increased spatial resolution down to the level of individual residues. A practical overview of how to couple ETD to the classical bottom-up HDX-MS workflow is given and new options for method optimization are discussed and exemplified. In addition, the real-world applicability of the HDX-ETD method to pinpoint conformational changes in a large 75 kDa protein complex of therapeutic interest is demonstrated. This feature highlights how the conformation and interactions of complex protein systems of biological or pharmaceutical interest can now be analyzed at a hitherto unprecedented level of structural detail using an MS-based method.

Figure optionsDownload high-quality image (163 K)Download as PowerPoint slideHighlights
► The feature details how electron transfer dissociation (ETD) can be used to measure protein hydrogen/deuterium exchange (HDX) at the level of individual residues.
► A practical overview of the experiment is given and new options for method optimization are discussed through the use of relevant examples.
► The applicability of the HDX-ETD method to pinpoint conformational changes in a large therapeutically relevant protein complex is demonstrated.
► The feature highlights how the conformation and interactions of complex proteins can be analyzed at a hitherto unprecedented level of detail using MS.