Clathrin senses membrane curvature

Zeno, Wade F.; Hochfelder, Jacob B.; Thatte, Ajay S.; Wang, Liping; Gadok, Avinash K.; Hayden, Carl C.; Lafer, Eileen M.; Stachowiak, Jeanne C.

doi:10.1016/j.bpj.2020.12.035

Cited by 37 publications

(22 citation statements)

References 49 publications

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“…Whilst considerable deformation is still required to form a vesicle at these pre-curved sites, the total energy requirement is lower than at a flat sheet, further indicating a clathrin-only mechanism is plausible. Use of nanofabricated surfaces has shown that inducing inward curvature on this scale promotes CCP formation at the plasma membrane (Zhao et al, 2017) and recent work suggests that clathrin prefers pre-curved surfaces (Zeno et al, 2021). Moreover in many cell lines, clathrin-coated pits often occur adjacent to natural plasma membrane protrusions where there is inward curvature, which may reflect a similar preference during endocytosis (Shevchuk et al, 2012); hinting that the clathrin may dominate membrane bending during endocytic vesicle formation.…”

Section: Discussionmentioning

confidence: 99%

Recruitment of clathrin to intracellular membranes is sufficient for vesicle formation

Küey¹,

Sittewelle²,

Larocque

et al. 2022

Preprint

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The formation of a clathrin-coated vesicle is a major membrane remodeling process that is crucial for membrane traffic in cells. Besides clathrin, these vesicles contain at least 100 different proteins although it is unclear how many are essential for the formation of the vesicle. Here, we show that intracellular clathrin-coated formation can be induced in living cells using minimal machinery and that it can be achieved on various membranes, including the mitochondrial outer membrane. Chemical heterodimerization was used to inducibly attach a clathrin-binding fragment "hook" to an "anchor" protein targeted to a specific membrane. Endogenous clathrin assembled to form coated pits on the mitochondria, termed MitoPits, within seconds of induction. MitoPits are double-membraned invaginations that form preferentially on high curvature regions of the mitochondrion. Upon induction, all stages of CCV formation - initiation, invagination, and even fission - were faithfully reconstituted. We found no evidence for the functional involvement of accessory proteins in this process. In addition, fission of MitoPit-derived vesicles was independent of known scission factors including dynamins and dynamin-related protein 1 (Drp1), suggesting that the clathrin cage generates sufficient force to bud intracellular vesicles. Our results suggest that, following its recruitment, clathrin alone is sufficient for intracellular clathrin-coated vesicle formation.

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Section: Discussionmentioning

confidence: 99%

Recruitment of clathrin to intracellular membranes is sufficient for vesicle formation

Küey¹,

Sittewelle²,

Larocque

et al. 2022

Preprint

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“…However, in solution cages are curved 51 , indicating that curved cages are more stable, absent the external constraints of the membrane. Recent experiments also quantified that clathrin assembly is more stable onto more highly curved surfaces, illustrating the energy gain that accompanies curved clathrin contacts 57 . Consistent with these observations, when we use the same energetic and kinetic parameters to simulate curved cage formation in solution, we see minimal assembly (Fig.…”

Section: Assembly In Solution Is Less Cooperative Than On the Membrane Despite Increased Stability Of The Curved Structuresmentioning

confidence: 99%

Nascent clathrin lattices spontaneously disassemble without sufficient adaptor proteins

Guo

Aj²,

Johnson

2021

Preprint

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Clathrin-coated structures must assemble on cell membranes to perform their primary function of receptor internalization. These structures show marked plasticity and instability, but what conditions are necessary to stabilize against disassembly have not been quantified. Recent in vitro fluorescence experiments have measured kinetics of stable clathrin assembly on membranes as controlled by key adaptor proteins like AP-2. Here, we combine this experimental data with microscopic reaction-diffusion simulations and theory to quantify mechanisms of stable vs unstable clathrin assembly on membranes. Both adaptor binding and dimensional reduction on the 2D surface are necessary to reproduce the cooperative kinetics of assembly. By applying our model to more physiologic-like conditions, where the stoichiometry and volume to area ratio are significantly lower than in vitro, we show that the critical nucleus contains ~25 clathrin, remarkably similar to sizes of abortive structures observed in vivo. Stable nucleation requires a stoichiometry of adaptor to clathrin that exceeds 1:1, meaning that AP-2 on its own has too few copies to nucleate lattices. Increasing adaptor concentration increases lattice sizes and nucleation speeds. For curved clathrin cages, we quantify both the cost of bending the membrane and the stabilization required to nucleate cages in solution. We find the energetics are comparable, suggesting that curving the lattice could offset the bending energy cost. Our model predicts how adaptor density controls stabilization of clathrin-coated structures against spontaneous disassembly, and shows remodeling and disassembly does not require ATPases, which is a critical advance towards predicting control of productive vesicle formation.

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“… 2019b ; Zeno et al. 2021 ). For example, full-length Amphiphysin1 (FL-Amph) and Epsin1 (FL-Epsin) sense membrane curvature more effectively than their structured domains, N-BAR and ENTH, respectively (Zeno et al.…”

Section: Idps/idprs-membrane Interactions: a Biophysical Studymentioning

confidence: 99%

“…Adapted from Zeno et al. ( 2018 , 2019a , 2019b , 2021 ) Protein/domain Domain types Membrane-binding domain Preferred lipid Sensing Induction FL-Espin OPD, IDPD ENTH AH, Entropic MC d His-Epsin a OPD, IDPD His-tag DGS-NTA Entropic MC wt-ENTH b OPD ENTH AH MC His- ENTH OPD His-tag DGS-NTA Insensitive MC His-EpsinCTD c IDPD His-tag DGS-NTA Entropic MC His-AP180CTD c IDPD His-tag DGS-NTA Entropic MC FL-Amph OPD, IDPD N-BAR AH, IC, Entropic AH, IC, OLG, MC Amph N-BAR OPD N-BAR A...…”

Section: Idps/idprs-membrane Interactions: a Biophysical Studymentioning

confidence: 99%

Insights into Membrane Curvature Sensing and Membrane Remodeling by Intrinsically Disordered Proteins and Protein Regions

2022

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Cellular membranes are highly dynamic in shape. They can rapidly and precisely regulate their shape to perform various cellular functions. The protein’s ability to sense membrane curvature is essential in various biological events such as cell signaling and membrane trafficking. As they are bound, these curvature-sensing proteins may also change the local membrane shape by one or more curvature driving mechanisms. Established curvature-sensing/driving mechanisms rely on proteins with specific structural features such as amphipathic helices and intrinsically curved shapes. However, the recent discovery and characterization of many proteins have shattered the protein structure–function paradigm, believing that the protein functions require a unique structural feature. Typically, such structure-independent functions are carried either entirely by intrinsically disordered proteins or hybrid proteins containing disordered regions and structured domains. It is becoming more apparent that disordered proteins and regions can be potent sensors/inducers of membrane curvatures. In this article, we outline the basic features of disordered proteins and regions, the motifs in such proteins that encode the function, membrane remodeling by disordered proteins and regions, and assays that may be employed to investigate curvature sensing and generation by ordered/disordered proteins. Graphical Abstract

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Clathrin senses membrane curvature

Cited by 37 publications

References 49 publications

Recruitment of clathrin to intracellular membranes is sufficient for vesicle formation

Recruitment of clathrin to intracellular membranes is sufficient for vesicle formation

Nascent clathrin lattices spontaneously disassemble without sufficient adaptor proteins

Insights into Membrane Curvature Sensing and Membrane Remodeling by Intrinsically Disordered Proteins and Protein Regions

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