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Cited by 52Cellular transport and secretionLipid Membrane Structure and BehaviorFungal and yeast genetics research

Epsin and Sla2 form assemblies through phospholipid interfaces

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Garcia-Alai, Maria M., Heidemann, Johannes, Skruzny, Michal, Gieras, Anna, Mertens, Haydyn D. T., Svergun, Dmitri I., Kaksonen, Marko, Uetrecht, Charlotte +1 more·2018·Nature Communications
DOI 10.1038/s41467-017-02443-xPMC PMC5780493
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Abstract

In clathrin-mediated endocytosis, adapter proteins assemble together with clathrin through interactions with specific lipids on the plasma membrane. However, the precise mechanism of adapter protein assembly at the cell membrane is still unknown. Here, we show that the membrane–proximal domains ENTH of epsin and ANTH of Sla2 form complexes through phosphatidylinositol 4,5-bisphosphate (PIP2) lipid interfaces. Native mass spectrometry reveals how ENTH and ANTH domains form assemblies by sharing PIP2 molecules. Furthermore, crystal structures of epsin Ent2 ENTH domain from S. cerevisiae in complex with PIP2 and Sla2 ANTH domain from C. thermophilum illustrate how allosteric phospholipid binding occurs. A comparison with human ENTH and ANTH domains reveal only the human ENTH domain can form a stable hexameric core in presence of PIP2, which could explain functional differences between fungal and human epsins. We propose a general phospholipid-driven multifaceted assembly mechanism tolerating different adapter protein compositions to induce endocytosis.

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  • After clathrin recruitment, PI(4,5)P2 facilitates membrane deformation; epsin binds PI(4,5)P2 and inserts an amphipathic helix, and BAR domain proteins also contribute.

    “After clathrin recruitment, PI(4,5)P2 facilitates membrane deformation. Epsin binds to PI(4,5)P2, localising epsin to the endocytic site where it inserts an amphipathic helix for membrane deformation [101,102]. Accessory proteins with BAR (domain named after three proteins: Bin, Amphiphysin and Rvs that share the domain) domains, which also bind PI(4,5)P2, also contribute to deformation of the”

    From Phosphatidylinositol(4,5)bisphosphate: diverse functions at the plasma membrane (2020) · cites this paper as [101]

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Research works printed but not linked: 47

  1. [1]McMahon HT Boucrot E Membrane curvature at a glance J. Cell Sci. 2015 128 1065 1070 10.1242/jcs.114454 25774051 PMC4359918 · DOI
  2. [2]Merrifield CJ Kaksonen M Endocytic accessory factors and regulation of clathrin-mediated endocytosis Cold Spring Harb. Perspect. Biol. 2014 6 a016733 10.1101/cshperspect.a016733 25280766 PMC4413230 · DOI
  3. [3]Ferguson SM De Camilli P Dynamin, a membrane-remodelling GTPase Nat. Rev. Mol. Cell Biol. 2012 13 75 88 22233676 PMC3519936
  4. [4]Kirchhausen T Owen D Harrison SC Molecular structure, function, and dynamics of clathrin-mediated membrane traffic Cold Spring Harb. Perspect. Biol. 2014 6 a016725 10.1101/cshperspect.a016725 24789820 PMC3996469 · DOI
  5. [5]Ford MGJ Curvature of clathrin-coated pits driven by epsin Nature 2002 419 361 366 10.1038/nature01020 12353027 · DOI · Paper page
  6. [6]Ford MG Simultaneous binding of PtdIns(4,5)P2 and clathrin by AP180 in the nucleation of clathrin lattices on membranes Science 2001 291 1051 1055 10.1126/science.291.5506.1051 11161218 · DOI
  7. [7]Jackson LP A large-scale conformational change couples membrane recruitment to cargo binding in the AP2 clathrin adaptor complex Cell 2010 141 1220 1229 10.1016/j.cell.2010.05.006 20603002 PMC3655264 · DOI
  8. [8]Itoh T Role of the ENTH domain in phosphatidylinositol-4,5-bisphosphate binding and endocytosis Science 2001 291 1047 1051 10.1126/science.291.5506.1047 11161217 · DOI · Paper page
  9. [10]Kozlov MM Mechanisms shaping cell membranes Curr. Opin. Cell Biol. 2014 29 53 60 10.1016/j.ceb.2014.03.006 24747171 PMC4180517 · DOI
  10. [11]Stachowiak JC Membrane bending by protein-protein crowding Nat. Cell Biol. 2012 14 944 949 10.1038/ncb2561 22902598 · DOI
  11. [12]Skruzny M Molecular basis for coupling the plasma membrane to the actin cytoskeleton during clathrin-mediated endocytosis Proc. Natl Acad. Sci. USA 2012 109 E2533 E2542 10.1073/pnas.1207011109 22927393 PMC3458359 · DOI
  12. [13]Messa M Epsin deficiency impairs endocytosis by stalling the actin-dependent invagination of endocytic clathrin-coated pits eLife 2014 3 e03311 10.7554/eLife.03311 25122462 PMC4161027 · DOI · Paper page
  13. [14]Skruzny M An organized co-assembly of clathrin adaptors is essential for endocytosis Dev. Cell 2015 33 150 162 10.1016/j.devcel.2015.02.023 25898165 · DOI
  14. [15]Boulant S Kural C Zeeh JC Ubelmann F Kirchhausen T Actin dynamics counteract membrane tension during clathrin-mediated endocytosis Nat. Cell Biol. 2011 13 1124 1131 10.1038/ncb2307 21841790 PMC3167020 · DOI
  15. [16]Erba EB Zenobi R Mass spectrometric studies of dissociation constants of noncovalent complexes Annu Rep. Prog. Chem. C Phys. Chem. 2011 107 199 228 10.1039/c1pc90006d · DOI
  16. [18]Sun J Kitova EN Wang W Klassen JS Method for distinguishing specific from nonspecific protein-ligand complexes in nanoelectrospray ionization mass spectrometry Anal. Chem. 2006 78 3010 3018 10.1021/ac0522005 16642987 · DOI · Paper page
  17. [19]Kitova EN El-Hawiet A Schnier PD Klassen JS Reliable determinations of protein–ligand interactions by direct esi-ms measurements. are we there yet? J. Am. Soc. Mass Spectrom. 2012 23 431 441 10.1007/s13361-011-0311-9 22270873 · DOI
  18. [20]Klotz IM Ligand–receptor interactions: facts and fantasies Q. Rev. Biophys. 1985 18 227 10.1017/S0033583500000354 3916127 · DOI
  19. [21]Brady RJ Damer CK Heuser JE O’Halloran TJ Regulation of Hip1r by epsin controls the temporal and spatial coupling of actin filaments to clathrin-coated pits J. Cell Sci. 2010 123 3652 3661 10.1242/jcs.066852 20923836 PMC2964106 · DOI
  20. [22]Aguilar RC Epsin N-terminal homology domains perform an essential function regulating Cdc42 through binding Cdc42 GTPase-activating proteins Proc. Natl Acad. Sci. USA 2006 103 4116 4121 10.1073/pnas.0510513103 16537494 PMC1449656 · DOI
  21. [23]Brady RJ Wen Y O’Halloran TJ The ENTH and C-terminal domains of Dictyostelium epsin cooperate to regulate the dynamic interaction with clathrin-coated pits J. Cell Sci. 2008 121 3433 3444 10.1242/jcs.032573 18827012 · DOI
  22. [24]Krissinel E Stock-based detection of protein oligomeric states in jsPISA Nucleic Acids Res. 2015 43 W314 W319 10.1093/nar/gkv314 25908787 PMC4489313 · DOI
  23. [25]Yoon Y Molecular basis of the potent membrane-remodeling activity of the epsin 1 N-terminal homology domain J. Biol. Chem. 2010 285 531 540 10.1074/jbc.M109.068015 19880963 PMC2804201 · DOI
  24. [27]Raucher D Phosphatidylinositol 4,5-Bisphosphate functions as a second messenger that regulates Cytoskeleton–Plasma membrane adhesion Cell 2000 100 221 228 10.1016/S0092-8674(00)81560-3 10660045 · DOI · Paper page
  25. [28]Gupta K The role of interfacial lipids in stabilizing membrane protein oligomers Nature 2017 541 421 424 10.1038/nature20820 28077870 PMC5501331 · DOI
  26. [29]Boucrot E Membrane fission is promoted by insertion of amphipathic helices and is restricted by crescent BAR domains Cell 2012 149 124 136 10.1016/j.cell.2012.01.047 22464325 PMC3465558 · DOI
  27. [30]Busch DJ Intrinsically disordered proteins drive membrane curvature Nat. Commun. 2015 6 7875 10.1038/ncomms8875 26204806 PMC4515776 · DOI
  28. [31]Bock T An integrated approach for genome annotation of the eukaryotic thermophile chaetomium thermophilum Nucleic Acids Res. 2014 42 13525 13533 10.1093/nar/gku1147 25398899 PMC4267624 · DOI
  29. [32]Bunkóczi G Phaser.MRage: automated molecular replacement Acta Crystallogr. D Biol. Crystallogr. 2013 69 2276 2286 10.1107/S0907444913022750 24189240 PMC3817702 · DOI
  30. [33]Winn MD Murshudov GN Papiz MZ Macromolecular TLS refinement in REFMAC at moderate resolutions Methods Enzymol. 2003 374 300 321 10.1016/S0076-6879(03)74014-2 14696379 · DOI
  31. [34]Emsley P Lohkamp B Scott WG Cowtan K Features and development of Coot Acta Crystallogr. D Biol. Crystallogr. 2010 66 486 501 10.1107/S0907444910007493 20383002 PMC2852313 · DOI
  32. [36]Chen VB MolProbity: all-atom structure validation for macromolecular crystallography Acta Crystallogr. D Biol. Crystallogr. 2010 66 12 21 10.1107/S0907444909042073 20057044 PMC2803126 · DOI · Paper page
  33. [37]Pettersen EF UCSF Chimera--a visualization system for exploratory research and analysis J. Comput. Chem. 2004 25 1605 1612 10.1002/jcc.20084 15264254 · DOI
  34. [38]Nurizzo D RoboDiff: combining a sample changer and goniometer for highly automated macromolecular crystallography experiments Acta Crystallogr. D Struct. Biol. 2016 72 966 975 10.1107/S205979831601158X 27487827 PMC4973212 · DOI
  35. [39]Kryshtafovych A Krysko O Daniluk P Dmytriv Z Fidelis K Protein structure prediction center in CASP8 Proteins 2009 77 5 9 10.1002/prot.22517 19722263 PMC2863353 · DOI
  36. [40]Langer GG Hazledine S Wiegels T Carolan C Lamzin VS Visual automated macromolecular model building Acta Crystallogr. D Biol. Crystallogr. 2013 69 635 641 10.1107/S0907444913000565 23519672 PMC3606041 · DOI
  37. [41]Blanchet CE Versatile sample environments and automation for biological solution X-ray scattering experiments at the P12 beamline (PETRA III, DESY) J. Appl. Crystallogr. 2015 48 431 443 10.1107/S160057671500254X 25844078 PMC4379436 · DOI
  38. [42]Petoukhov MV New developments in the ATSAS program package for small-angle scattering data analysis J. Appl. Crystallogr. 2012 45 342 350 10.1107/S0021889812007662 25484842 PMC4233345 · DOI
  39. [43]Semenyuk AV Svergun DI GNOM – a program package for small-angle scattering data processing J. Appl. Crystallogr. 1991 24 537 540 10.1107/S002188989100081X · DOI
  40. [44]Franke D Svergun DI DAMMIF, a program for rapid ab-initio shape determination in small-angle scattering J. Appl. Crystallogr. 2009 42 342 346 10.1107/S0021889809000338 27630371 PMC5023043 · DOI · Paper page
  41. [45]Lorenzen, K. & van Duijn, E. in Current Protocols in Protein Science (eds Coligan, J. E., Dunn, B. M., Speicher, D. W. & Wingfield, P. T.) 17.12.1–17.12.17 (John Wiley & Sons, Inc., New York, 2010).
  42. [46]Gamper N Shapiro MS Target-specific PIP 2 signalling: how might it work?: Target-specific PIP 2 signalling J. Physiol. 2007 582 967 975 10.1113/jphysiol.2007.132787 17412762 PMC2075238 · DOI · Paper page
  43. [47]van den Heuvel RHH Improving the performance of a quadrupole time-of-flight instrument for macromolecular mass spectrometry Anal. Chem. 2006 78 7473 7483 10.1021/ac061039a 17073415 · DOI
  44. [48]Lorenzen K Versluis C van Duijn E van den Heuvel RHH Heck AJR Optimizing macromolecular tandem mass spectrometry of large non-covalent complexes using heavy collision gases Int. J. Mass Spectrom. 2007 268 198 206 10.1016/j.ijms.2007.06.012 · DOI
  45. [49]Tahallah N Pinkse M Maier CS Heck AJR The effect of the source pressure on the abundance of ions of noncovalent protein assemblies in an electrospray ionization orthogonal time-of-flight instrument Rapid Commun. Mass Spectrom. 2001 15 596 601 10.1002/rcm.275 11312509 · DOI
  46. [50]Morgner N Robinson CV Mass ign: an assignment strategy for maximizing information from the mass spectra of heterogeneous protein assemblies Anal. Chem. 2012 84 2939 2948 10.1021/ac300056a 22409725 · DOI
  47. [51]Sun Y Carroll S Kaksonen M Toshima JY Drubin DG PtdIns(4,5)P2 turnover is required for multiple stages during clathrin- and actin-dependent endocytic internalization J. Cell Biol. 2007 177 355 367 10.1083/jcb.200611011 17452534 PMC2064142 · DOI

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Authors
Garcia-Alai, M. M., Heidemann, J., Skruzny, M., Gieras, A., Mertens, H. D. T., Svergun, D. I., Kaksonen, M., Uetrecht, C., & Meijers, R.
Affiliations
  • Leibniz Institute of Virology (LIV) (DE)
  • University Medical Center Hamburg-Eppendorf (DE)
  • European Molecular Biology Laboratory (DE)
  • University of Geneva (CH)
Corresponding author
  • Uetrecht, Charlotte
  • Meijers, Rob
ORCID
Venue
Nature Communications
Publisher
Springer Science and Business Media LLC
Published
2018-01-23
Volume
9
Issue
1
Pages
328-328
ISSN
2041-1723 (ISSN-L)
DOI
10.1038/s41467-017-02443-x
PMC
PMC5780493
OpenAlex
W2793872756
Cited by
52
References
4
Citations by year
2026: 1 · 2025: 3 · 2024: 5 · 2023: 9 · 2022: 7 · 2021: 8 · 2020: 11 · 2019: 6 · 2018: 2
FWCI
3.74
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Communications (publishedVersion, cc-by)
  • Europe PMC (PubMed Central) (publishedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Archive ouverte UNIGE (University of Geneva) (publishedVersion, other-oa)
  • RePEc: Research Papers in Economics (submittedVersion)
  • Desy Publications Database (Deutsches Elektronen-Synchrotron DESY) (submittedVersion, other-oa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
  • MPG.PuRe (Max Planck Society) (submittedVersion)
  • Archive ouverte UNIGE (University of Geneva) (submittedVersion, cc-by)
  • European XFEL (submittedVersion)
  • DESY Publication Database (PUBDB) (Deutsches Elektronen-Synchrotron)
Funders
  • National Science Foundation
  • European Synchrotron Radiation Facility
  • Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung (163267)
  • Bundesministerium für Gesundheit
  • Deutsches Elektronen-Synchrotron
  • Leibniz-Gemeinschaft (SAW‐2014‐HPI‐4)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular transport and secretion
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Lipid Membrane Structure and Behavior
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Fungal and yeast genetics research
Keywords
epsin, Sla2, clathrin-mediated endocytosis, phosphatidylinositol 4,5-bisphosphate, ENTH domain, thermophilic archaea
MeSH terms
Protein Domains, Amino Acid Sequence, Binding Sites, Cell Membrane, Chaetomium, Endocytosis, Fungal Proteins, Humans, Models, Molecular, Protein Binding, Saccharomyces cerevisiae, Sequence Homology, Amino Acid, Crystallography, X-Ray, Phosphatidylinositol 4,5-Diphosphate, Adaptor Proteins, Vesicular Transport, Protein Multimerization

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