Efficient Protein Digestion at Elevated Temperature in the Presence of SDS and Calcium Ions for Membrane Proteomics.

Growing significance of membrane proteins inspires continuous development and improvement of methods for robust membrane proteomics. Here, we developed a very simple and efficient method for membrane protein digestion using an ionic detergent SDS at high temperature, conditions where trypsin is normally inactivated. Our results suggest that trypsin can be stabilized by a combination of calcium ions and sodium chloride which enables protein digestion at elevated temperature in the presence of SDS. Application of this method resulted in higher efficiency of digestion and improved identification of membrane proteins, offering novel opportunities for advancement in membrane proteomics.