Abstract
Specific post-translational modification (PTM) states of a protein affect its property and function; understanding their dynamics in cells would provide deep insight into diverse signaling pathways and biological processes. However, it is not trivial to visualize post-translational modifications in a protein- and site-specific manner, especially in a living-cell context. Herein, we review recent advances in the development of molecular imaging tools to detect diverse classes of post-translational proteoforms in individual cells, and their applications in studying precise roles of PTMs in regulating the function of cellular proteins.
Highlights
Linking a precise protein form to its property and function in the crowded cellular context remains an outstanding challenge in biological research
post-translational modification (PTM) dynamics can be tracked in real time via monitoring the subcellular activity of post-translational modifying enzymes using a suite of Forster resonance energy transfer (FRET)-based biosensors containing surrogate substrates for the enzymes; efforts to create this class of technologies to probe diverse biological systems have been reviewed.[30]
We discuss platforms for high-throughput and multiplexed protein imaging, which can be readily applied to imaging of cellular proteoforms, once suitable molecular probes for proteoform detection are in place
Summary
Of proteoforms is often not retained in mass spectrometry-based proteomic studies unless the investigators employ cellular fractionation[11] or proximity labeling strategies[12] to selectively enrich proteomes from a given location within the cell These technologies enable global and subcellular profiling of bona fide proteoforms for further functional characterizations. PTM dynamics can be tracked in real time via monitoring the subcellular activity of post-translational modifying enzymes (kinases/phosphatases, methyltransferases, and glycosyltransferases) using a suite of Forster resonance energy transfer (FRET)-based biosensors containing surrogate substrates for the enzymes; efforts to create this class of technologies to probe diverse biological systems have been reviewed.[30] complementary molecular tools which enable direct visualization of PTM dynamics on desired protein targets are not as welldeveloped. We discuss platforms for high-throughput and multiplexed protein imaging, which can be readily applied to imaging of cellular proteoforms, once suitable molecular probes for proteoform detection are in place
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