The dynamic proteome: Expanding the toolkit for translation proteomics
Cells can adapt rapidly to changing environments. These responses are often shaped by alterations in the translational landscape of the cell. Quantifying these changes in an unbiased way provides with new insights to translational regulation and molecular mechanisms. To facilitate faithful translation quantification and to overcome technical limitations we developed multiplexed enhanced Protein Dynamics (mePROD) mass spectrometry. The combination of Tandem Mass Tags and SILAC pulse labeling together with a carrier or booster signal enables us to profile translatome wide changes in the range of minutes without loosing depth of anaylsis. mePROD is able to accurately measure the translation of thousands of proteins, even under conditions where global translation rates are effected (e.g. stress responses). We successfully appied mePROD to diverse disease scenarios, such as viral infection. We currently integrating functional translation proteomics into a wide range of applications to spark new ideas in disease biology.
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A detailed protocol can be found here:
Klann and Münch, Methods Mol Biol 2022,2428:75-87.