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Size-dependent protein segregation at membrane interfaces


Membrane interfaces formed at cell–cell junctions are associated with characteristic patterns of membrane proteins whose organization is critical for intracellular signalling. To isolate the role of membrane protein size in pattern formation, we reconstituted model membrane interfaces in vitro using giant unilamellar vesicles decorated with synthetic binding and non-binding proteins. We show that size differences between membrane proteins can drastically alter their organization at membrane interfaces, with as little as a 5 nm increase in non-binding protein size driving its exclusion from the interface. Combining in vitro measurements with Monte Carlo simulations, we find that non-binding protein exclusion is also influenced by lateral crowding, binding protein affinity, and thermally driven membrane height fluctuations that transiently limit access to the interface. This sensitive and highly effective means of physically segregating proteins has implications for cell–cell contacts such as T-cell immunological synapses (for example, CD45 exclusion) and epithelial cell junctions (for example, E-cadherin enrichment), as well as for protein sorting at intracellular contact points between membrane-bound organelles.

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Figure 1: In vitro membrane interface system.
Figure 2: Effect of non-binding protein height on segregation at membrane interfaces.
Figure 3: Membrane bending by long non-binding proteins at membrane interfaces.
Figure 4: Effect of protein crowding on segregation at membrane interfaces.
Figure 5: Monte Carlo simulations of size-dependent protein segregation.


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We acknowledge R. Vale and C. Peel for helpful discussions. This work was supported by a Graduate Fellows Research Program grant from the National Science Foundation (NSF) for M.H.B.; a Cancer Research Institute Post-Doctoral Fellowship and a K99 grant from the National Institute of Health (NIH, K99AI093884 and R00AI093884) for K.C.; a Forschungsstipendium of the Deutsche Forschungsgemeinschaft (DFG grant no. We 5004/2) for J.W.; a NIH grant (R37AI043542), a NIGMS Nanomedicine Development Center grant (PN2EY016586) and a Wellcome Trust Principal Research Fellowship to M.L.D.; and a NIH Nanomedicine Development Center grant (PN2EY016546) and an NIH R01 grant (GM114344) to D.A.F. This research was also supported by the Chemical Sciences, Geosciences and Biosciences Division, Office of Basic Energy Sciences, Office of Science, US Department of Energy, FWP number SISGRKN.

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All authors contributed to design of the experiments. E.M.S. and M.H.B. performed experiments. E.M.S., H.S.A. and M.H.B. created unique materials. J.W., M.H.B. and P.L.G. performed simulations and modelling. E.M.S., M.H.B. and D.A.F. wrote the manuscript. All authors discussed the results and commented on the manuscript.

Corresponding authors

Correspondence to Eva M. Schmid or Matthew H. Bakalar or Daniel A. Fletcher.

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The authors declare no competing financial interests.

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Schmid, E., Bakalar, M., Choudhuri, K. et al. Size-dependent protein segregation at membrane interfaces. Nature Phys 12, 704–711 (2016).

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