An endophilin–dynamin complex promotes budding of clathrin-coated vesicles during synaptic vesicle recycling

Sundborger, Anna; Soderblom, Cynthia; Vorontsova, Olga; Evergren, Emma; Hinshaw, Jenny E.; Shupliakov, Oleg

doi:10.1242/jcs.072686

Cited by 116 publications

(121 citation statements)

References 35 publications

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“…Dynasore was used to disrupt CME. This pharmacological reagent works by inhibiting the GTPase dynamin (22), which is mainly responsible for the scission of vesicular buds from the plasma membrane (34). It has been suggested that dynamin is also involved in the early stages of CCP formation, serving as both a regulator and integrity monitor (10,35,36).…”

Section: Methodsmentioning

confidence: 99%

Quantifying the dynamic interactions between a clathrin-coated pit and cargo molecules

Weigel¹,

Tamkun²,

Krapf

2013

Proc. Natl. Acad. Sci. U.S.A.

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Clathrin-mediated endocytosis takes place through the recruitment of cargo molecules into a growing clathrin-coated pit (CCP). Despite the importance of this process to all mammalian cells, little is yet known about the interaction dynamics between cargo and CCPs. These interactions are difficult to study because CCPs display a large degree of lifetime heterogeneity and the interactions with cargo molecules are time dependent. We use single-molecule total internal reflection fluorescence microscopy, in combination with automatic detection and tracking algorithms, to directly visualize the recruitment of individual voltage-gated potassium channels into forming CCPs in living cells. We observe association and dissociation of individual channels with a CCP and, occasionally, their internalization. Contrary to widespread ideas, cargo often escapes from a pit before abortive CCP termination or endocytic vesicle production. Thus, the binding times of cargo molecules associating to CCPs are much shorter than the overall endocytic process. By measuring tens of thousands of capturing events, we build the distribution of capture times and the times that cargo remains confined to a CCP. An analytical stochastic model is developed and compared with the measured distributions. Due to the dynamic nature of the pit, the model is non-Markovian and it displays long-tail power law statistics. The measured distributions and model predictions are in excellent agreement over more than five orders of magnitude. Our findings identify one source of the large heterogeneities in CCP dynamics and provide a mechanism for the anomalous diffusion of proteins in the plasma membrane.single-molecule tracking | Levy statistics | live-cell imaging | TIRF

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Section: Methodsmentioning

confidence: 99%

Quantifying the dynamic interactions between a clathrin-coated pit and cargo molecules

Weigel¹,

Tamkun²,

Krapf

2013

Proc. Natl. Acad. Sci. U.S.A.

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“…The cluster-associated protein pool can be mobilized by synaptic activity. Thus, after a period of repetitive stimulation the endocytic proteins partly dissociate from the synaptic vesicles and appear at the plasma membrane in the periactive zone (30)(31)(32). Such protein redistibution has also been observed in live imaging experiments.…”

Section: A Storage Pool Of Endocytic Proteins Is Associated With Synamentioning

confidence: 80%

Key Events in Synaptic Vesicle Endocytosis

Ackermann¹,

Gregory²,

Brodin³

2012

Molecular Regulation of Endocytosis

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“…labeled with mCherry should form a complex in the cytoplasm (Ross et al, 2011;Sundborger et al, 2010). Endophilin in the cytoplasm has been found to be a dimer (Ringstad, Nemoto, & De Camilli, 2001;Ross et al, 2011).…”

Section: Applications Of Rics and Nandb To Detect Molecular Complexes Imentioning

confidence: 99%

Raster Image Correlation Spectroscopy and Number and Brightness Analysis

Digman¹,

Stakic²,

Gratton³

2013

Methods in Enzymology

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An endophilin–dynamin complex promotes budding of clathrin-coated vesicles during synaptic vesicle recycling

Cited by 116 publications

References 35 publications

Quantifying the dynamic interactions between a clathrin-coated pit and cargo molecules

Quantifying the dynamic interactions between a clathrin-coated pit and cargo molecules

Key Events in Synaptic Vesicle Endocytosis

Raster Image Correlation Spectroscopy and Number and Brightness Analysis

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