Temporal Tracking of Insulin Action on the Cell Surface of Proteins at a Resolution of Ten Seconds

The ligand–receptor signaling occurring on the
cell surface
governs cell growth, proliferation, and survival via rapidly triggering
a cascade of events. Here, we for the first time report an in situ perturbation-free and rapid surface proteomic profiling
at a temporal resolution of ten seconds. By this innovation, about
1022 cell surface-associated proteins were reproducibly identified
and quantified. It is noteworthy that, upon a model ligand insulin
stimulus, a few rapid-responding proteins at 10 s to 2 min were identified,
e.g., CNNM3. Moreover, temporal response patterns were established
for the members of GLUT4 storage vesicles (GSVs; responsible for glucose
transportation) and confirmed with five known GSV proteins. This pattern
was then exploited to uncover seven new regulatory proteins (LDLR,
HFE, ECE1, MRC2, CORO1C, CPD, and BST2). Collectively, we showed a
powerful surface proteomic tool to decipher rapid signaling of cell-surface
proteins and to uncover new subunits involved in rapidly trafficking
vesicles.