A High Precision Survey of the Molecular Dynamics of Mammalian Clathrin-Mediated Endocytosis

Abstract

The molecular dynamics of clathrin-mediated endocytosis in living cells has been mapped with an approximately ten-fold improvement in temporal accuracy, yielding new insights into the molecular mechanism.

Key Events in Synaptic Vesicle Endocytosis

“…Dynamin is present at low levels from early stages but exhibits a peak just before scission. A similar behavior is also observed for endophilin and synaptojanin (38,39).…”

“…This contrasts with the detailed information that has been obtained in non-neuronal cells grown on glass slides in which protein movement near the plasma membrane can be tracked by total internal reflection (TIRF) microcopy (38). These studies indicate that among the first proteins to occur at the plasma membran is the F-BAR protein FCHo1/2, followed by the scaffold proteins eps15 and intersectin.…”

pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis

“…The ppH protocol has been extensively used for this purpose (Taylor et al, 2011). However, the need to use the green channel for the ± 1.7% of clusters detected with clc-GFP or TfR-pHuji, respectively, contain both markers (Fig.…”

“…For example, a red FP pH sensor could be imaged in conjunction with a GFP-tagged protein of interest to analyze the spatial and temporal dynamics of protein recruitment during exocytosis or endocytosis (Taylor et al, 2011). Alternatively, the trafficking of two membrane cargos could be monitored simultaneously to address their differential sorting.…”

Linking up at the BAR: Oligomerization and F-BAR protein function

“…53 Both FBP17 and CIP4's SH3 domains also recruit the GTPase Dynamin, 6,21,28,50 a critical component for vesicle scission 54 ( Fig. 2).…”