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bioRxiv · 10.1101/2024.07.23.604744

Site-specific metabolic labeling reveals proteome-wide phospho-dynamics

Alevra, M.·Correa Marrero, M.·Kluever, V.·Mandad, S.·Hemandhar-Kumar, N.·Pan, K.-T.·van Gerwen, J.·Glowacki, S. K.·Wildhagen, H.·Ischebeck, T.·Liu, Y.·RIzzoli, S. O.·Urlaub, H.·Beltrao, P.·Fornasiero, E. F.
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Abstract

Protein phosphorylation is a critical post-translational modification that orchestrates cellular signaling. Here, we introduce PulsPhos, a method combining metabolic labeling with phosphoproteomics, spectral analysis and modeling, to quantify site-specific phosphorylation lifetimes in living cells. Phosphosite lifetimes vary over multiple orders of magnitude and are influenced by factors such as amino acid composition and subcellular localization. PulsPhos was readily applied to pharmacological perturbations revealing fundamental mechanisms governing protein phosphorylation dynamics.

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Alevra, M., Correa Marrero, M., Kluever, V., Mandad, S., Hemandhar-Kumar, N., Pan, K.-T., van Gerwen, J., Glowacki, S. K., Wildhagen, H., Ischebeck, T., Liu, Y., RIzzoli, S. O., Urlaub, H., Beltrao, P., Fornasiero, E. F.. 2024-07-24. Site-specific metabolic labeling reveals proteome-wide phospho-dynamics. https://doi.org/10.1101/2024.07.23.604744

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