Cavin1 intrinsically disordered domains are essential for fuzzy electrostatic interactions and caveola formation.
Journal
Nature communications
ISSN: 2041-1723
Titre abrégé: Nat Commun
Pays: England
ID NLM: 101528555
Informations de publication
Date de publication:
10 02 2021
10 02 2021
Historique:
received:
16
09
2020
accepted:
06
01
2021
entrez:
11
2
2021
pubmed:
12
2
2021
medline:
19
3
2021
Statut:
epublish
Résumé
Caveolae are spherically shaped nanodomains of the plasma membrane, generated by cooperative assembly of caveolin and cavin proteins. Cavins are cytosolic peripheral membrane proteins with negatively charged intrinsically disordered regions that flank positively charged α-helical regions. Here, we show that the three disordered domains of Cavin1 are essential for caveola formation and dynamic trafficking of caveolae. Electrostatic interactions between disordered regions and α-helical regions promote liquid-liquid phase separation behaviour of Cavin1 in vitro, assembly of Cavin1 oligomers in solution, generation of membrane curvature, association with caveolin-1, and Cavin1 recruitment to caveolae in cells. Removal of the first disordered region causes irreversible gel formation in vitro and results in aberrant caveola trafficking through the endosomal system. We propose a model for caveola assembly whereby fuzzy electrostatic interactions between Cavin1 and caveolin-1 proteins, combined with membrane lipid interactions, are required to generate membrane curvature and a metastable caveola coat.
Identifiants
pubmed: 33568658
doi: 10.1038/s41467-021-21035-4
pii: 10.1038/s41467-021-21035-4
pmc: PMC7875971
doi:
Substances chimiques
Caveolin 1
0
Cavin1 protein, mouse
0
Membrane Proteins
0
RNA-Binding Proteins
0
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
931Références
Lamaze, C., Tardif, N., Dewulf, M., Vassilopoulos, S. & Blouin, C. M. The caveolae dress code: structure and signaling. Curr. Opin. Cell Biol. 47, 117–125 (2017).
pubmed: 28641181
doi: 10.1016/j.ceb.2017.02.014
Parton, R. G. & del Pozo, M. A. Caveolae as plasma membrane sensors, protectors and organizers. Nat. Rev. Mol. Cell Biol. 14, 98–112 (2013).
pubmed: 23340574
doi: 10.1038/nrm3512
Parton, R. G. Caveolae: structure, function, and relationship to disease. Annu. Rev. Cell Dev. Biol. 34, 111–136 (2018).
pubmed: 30296391
doi: 10.1146/annurev-cellbio-100617-062737
Hayer, A., Stoeber, M., Bissig, C. & Helenius, A. Biogenesis of caveolae: stepwise assembly of large caveolin and cavin complexes. Traffic 11, 361–382 (2010).
pubmed: 20070607
doi: 10.1111/j.1600-0854.2009.01023.x
Ariotti, N. et al. Molecular characterization of caveolin-induced membrane curvature. J. Biol. Chem. 290, 24875–24890 (2015).
pubmed: 26304117
pmcid: 4598997
doi: 10.1074/jbc.M115.644336
Busija, A. R., Patel, H. H. & Insel, P. A. Caveolins and cavins in the trafficking, maturation, and degradation of caveolae: implications for cell physiology. Am. J. Physiol. Cell Physiol. 312, C459–C477 (2017).
pubmed: 28122734
pmcid: 5407024
doi: 10.1152/ajpcell.00355.2016
Hill, M. M. et al. PTRF-cavin, a conserved cytoplasmic protein required for caveola formation and function. Cell 132, 113–124 (2008).
pubmed: 18191225
pmcid: 2265257
doi: 10.1016/j.cell.2007.11.042
Walser, P. J. et al. Constitutive formation of caveolae in a bacterium. Cell 150, 752–763 (2012).
pubmed: 22901807
doi: 10.1016/j.cell.2012.06.042
Bastiani, M. et al. MURC/Cavin-4 and cavin family members form tissue-specific caveolar complexes. J. Cell Biol. 185, 1259–1273 (2009).
pubmed: 19546242
pmcid: 2712963
doi: 10.1083/jcb.200903053
Tillu, V. A. et al. A variable undecad repeat domain in cavin1 regulates caveola formation and stability. EMBO Rep. 19, e45775 (2018).
pubmed: 30021837
pmcid: 6123655
doi: 10.15252/embr.201845775
Kovtun, O. et al. Structural insights into the organization of the cavin membrane coat complex. Dev. Cell 31, 405–419 (2014).
pubmed: 25453557
doi: 10.1016/j.devcel.2014.10.002
Kovtun, O., Tillu, V. A., Ariotti, N., Parton, R. G. & Collins, B. M. Cavin family proteins and the assembly of caveolae. J. Cell Sci. 128, 1269–1278 (2015).
pubmed: 25829513
pmcid: 4379724
doi: 10.1242/jcs.167866
Mohan, J., Moren, B., Larsson, E., Holst, M. & Lundmark, R. Cavin3 interacts with cavin1 and caveolin1 to increase surface dynamics of caveolae. J. Cell Sci. 128, 979–991 (2015).
pubmed: 25588833
Gambin, Y. et al. Single-molecule analysis reveals self assembly and nanoscale segregation of two distinct cavin subcomplexes on caveolae. eLife 3, e01434 (2014).
pmcid: 3903133
doi: 10.7554/eLife.01434
Oates, M. E. et al. D2P2: database of disordered protein predictions. Nucleic Acids Res. 41, D508–D516 (2012).
pubmed: 23203878
pmcid: 3531159
doi: 10.1093/nar/gks1226
Banks, D. S. & Fradin, C. Anomalous diffusion of proteins due to molecular crowding. Biophys. J. 89, 2960–2971 (2005).
pubmed: 16113107
pmcid: 1366794
doi: 10.1529/biophysj.104.051078
Lavie, Y., Fiucci, G. & Liscovitch, M. Up-regulation of caveolae and caveolar constituents in multidrug-resistant cancer cells. J. Biol. Chem. 273, 32380–32383 (1998).
pubmed: 9829965
doi: 10.1074/jbc.273.49.32380
Tillu, V. A., Kovtun, O., McMahon, K. A., Collins, B. M. & Parton, R. G. A phosphoinositide-binding cluster in cavin1 acts as a molecular sensor for cavin1 degradation. Mol. Biol. Cell 26, 3561–3569 (2015).
pubmed: 26269585
pmcid: 4603927
doi: 10.1091/mbc.E15-06-0359
Boeynaems, S. et al. Protein phase separation: a new phase in cell biology. Trends Cell Biol. 28, 420–435 (2018).
pubmed: 29602697
pmcid: 6034118
doi: 10.1016/j.tcb.2018.02.004
Boyko, S., Qi, X., Chen, T. H., Surewicz, K. & Surewicz, W. K. Liquid–liquid phase separation of tau protein: the crucial role of electrostatic interactions. J. Biol. Chem. 294, 11054–11059 (2019).
pubmed: 31097543
pmcid: 6643045
doi: 10.1074/jbc.AC119.009198
Gomes, E. & Shorter, J. The molecular language of membraneless organelles. J. Biol. Chem. 294, 7115–7127 (2019).
pubmed: 30045872
doi: 10.1074/jbc.TM118.001192
Wang, J. et al. A molecular grammar governing the driving forces for phase separation of prion-like RNA binding proteins. Cell 174, 688–699 (2018).
pubmed: 29961577
pmcid: 6063760
doi: 10.1016/j.cell.2018.06.006
Alenquer, M. et al. Influenza A virus ribonucleoproteins form liquid organelles at endoplasmic reticulum exit sites. Nat. Commun. 10, 1629 (2019).
pubmed: 30967547
pmcid: 6456594
doi: 10.1038/s41467-019-09549-4
Case, L. B., Zhang, X., Ditlev, J. A. & Rosen, M. K. Stoichiometry controls activity of phase-separated clusters of actin signaling proteins. Science 363, 1093–1097 (2019).
pubmed: 30846599
pmcid: 6784323
doi: 10.1126/science.aau6313
Ditlev, J. A. et al. A composition-dependent molecular clutch between T cell signaling condensates and actin. eLife 8, e42695 (2019).
pubmed: 31268421
pmcid: 6624021
doi: 10.7554/eLife.42695
Huang, W. Y. C. et al. A molecular assembly phase transition and kinetic proofreading modulate Ras activation by SOS. Science 363, 1098–1103 (2019).
pubmed: 30846600
pmcid: 6563836
doi: 10.1126/science.aau5721
Johnson, A. et al. TFG clusters COPII-coated transport carriers and promotes early secretory pathway organization. EMBO J. 34, 811–827 (2015).
pubmed: 25586378
pmcid: 4369316
doi: 10.15252/embj.201489032
Liao, Y. C. et al. RNA granules Hitchhike on lysosomes for long-distance transport, using annexin A11 as a molecular tether. Cell 179, 147–164 (2019).
pubmed: 31539493
pmcid: 6890474
doi: 10.1016/j.cell.2019.08.050
Ma, W. & Mayr, C. A membraneless organelle associated with the endoplasmic reticulum enables 3′UTR-mediated protein–protein interactions. Cell 175, 1492–1506 e1419 (2018).
pubmed: 30449617
pmcid: 6711188
doi: 10.1016/j.cell.2018.10.007
Milovanovic, D., Wu, Y., Bian, X. & De Camilli, P. A liquid phase of synapsin and lipid vesicles. Science 361, 604–607 (2018).
pubmed: 29976799
pmcid: 6191856
doi: 10.1126/science.aat5671
Snead, W. T. & Gladfelter, A. S. The control centers of biomolecular phase separation: how membrane surfaces, PTMs, and active processes regulate condensation. Mol. Cell 76, 295–305 (2019).
pubmed: 31604601
pmcid: 7173186
doi: 10.1016/j.molcel.2019.09.016
Zappa, F. et al. The TRAPP complex mediates secretion arrest induced by stress granule assembly. EMBO J. 38, e101704 (2019).
pubmed: 31429971
pmcid: 6769382
doi: 10.15252/embj.2019101704
Beutel, O., Maraspini, R., Pombo-García, K., Martin-Lemaitre, C. & Honigmann, A. Phase separation of Zonula Occludens proteins drives formation of tight junctions. Cell 179, 923–936.e911 (2019).
pubmed: 31675499
doi: 10.1016/j.cell.2019.10.011
Schwayer, C. et al. Mechanosensation of tight junctions depends on ZO-1 phase separation and flow. Cell 179, 937–952.e918 (2019).
pubmed: 31675500
doi: 10.1016/j.cell.2019.10.006
Bergeron-Sandoval, L. P. & Michnick, S. W. Mechanics, structure and function of biopolymer condensates. J. Mol. Biol. 430, 4754–4761 (2018).
pubmed: 29913159
doi: 10.1016/j.jmb.2018.06.023
Lacy, M. M., Ma, R., Ravindra, N. G. & Berro, J. Molecular mechanisms of force production in clathrin-mediated endocytosis. FEBS Lett. 592, 3586–3605 (2018).
pubmed: 30006986
pmcid: 6231980
doi: 10.1002/1873-3468.13192
Bergeron-Sandoval, L.-P. et al. Endocytosis caused by liquid–liquid phase separation of proteins. Preprint at bioRxiv 145664 (2018).
Li, Y., Lipowsky, R. & Dimova, R. Membrane nanotubes induced by aqueous phase separation and stabilized by spontaneous curvature. Proc. Natl Acad. Sci. USA 108, 4731–4736 (2011).
pubmed: 21383120
doi: 10.1073/pnas.1015892108
pmcid: 3064332
Alberti, S. et al. A User’s guide for phase separation assays with purified proteins. J. Mol. Biol. 430, 4806–4820 (2018).
pubmed: 29944854
pmcid: 6215329
doi: 10.1016/j.jmb.2018.06.038
Wang, Z., Zhang, G. & Zhang, H. Protocol for analyzing protein liquid–liquid phase separation. Biophys. Rep. 5, 1–9 (2019).
doi: 10.1007/s41048-018-0078-7
Itzhak, D. N., Tyanova, S., Cox, J. & Borner, G. H. Global, quantitative and dynamic mapping of protein subcellular localization. eLife 5, e16950 (2016).
pubmed: 27278775
pmcid: 4959882
doi: 10.7554/eLife.16950
Molliex, A. et al. Phase separation by low complexity domains promotes stress granule assembly and drives pathological fibrillization. Cell 163, 123–133 (2015).
pubmed: 26406374
pmcid: 5149108
doi: 10.1016/j.cell.2015.09.015
Alberti, S., Gladfelter, A. & Mittag, T. Considerations and challenges in studying liquid–liquid phase separation and biomolecular condensates. Cell 176, 419–434 (2019).
pubmed: 30682370
pmcid: 6445271
doi: 10.1016/j.cell.2018.12.035
Kubala, M. H., Kovtun, O., Alexandrov, K. & Collins, B. M. Structural and thermodynamic analysis of the GFP:GFP-nanobody complex. Protein Sci. 19, 2389–2401 (2010).
pubmed: 20945358
pmcid: 3009406
doi: 10.1002/pro.519
Dewulf, M. et al. Dystrophy-associated caveolin-3 mutations reveal that caveolae couple IL6/STAT3 signaling with mechanosensing in human muscle cells. Nat. Commun. 10, 1974 (2019).
pubmed: 31036801
pmcid: 6488599
doi: 10.1038/s41467-019-09405-5
Hayer, A. et al. Caveolin-1 is ubiquitinated and targeted to intralumenal vesicles in endolysosomes for degradation. J. Cell Biol. 191, 615–629 (2010).
pubmed: 21041450
pmcid: 3003328
doi: 10.1083/jcb.201003086
Hernandez-Deviez, D. J. et al. Aberrant dysferlin trafficking in cells lacking caveolin or expressing dystrophy mutants of caveolin-3. Hum. Mol. Genet. 15, 129–142 (2006).
pubmed: 16319126
doi: 10.1093/hmg/ddi434
Sun, X. H. et al. A conserved sequence in caveolin-1 is both necessary and sufficient for caveolin polarity and cell directional migration. FEBS Lett. 583, 3681–3689 (2009).
pubmed: 19854179
doi: 10.1016/j.febslet.2009.10.055
Kirkham, M. et al. Evolutionary analysis and molecular dissection of caveola biogenesis. J. Cell Sci. 121, 2075–2086 (2008).
pubmed: 18505796
doi: 10.1242/jcs.024588
Sun, X. H. et al. Identification of a novel domain at the N terminus of caveolin-1 that controls rear polarization of the protein and caveolae formation. J. Biol. Chem. 282, 7232–7241 (2007).
pubmed: 17213184
doi: 10.1074/jbc.M607396200
Ingelmo-Torres, M. et al. Hydrophobic and basic domains target proteins to lipid droplets. Traffic 10, 1785–1801 (2009).
pubmed: 19874557
pmcid: 2913680
doi: 10.1111/j.1600-0854.2009.00994.x
Rothberg, K. G. et al. Caveolin, a protein component of caveolae membrane coats. Cell 68, 673–682 (1992).
doi: 10.1016/0092-8674(92)90143-Z
pubmed: 1739974
Shaul, P. W. & Anderson, R. G. Role of plasmalemmal caveolae in signal transduction. Am. J. Physiol. 275, L843–L851 (1998).
pubmed: 9815100
Ludwig, A. et al. Molecular composition and ultrastructure of the caveolar coat complex. PLoS Biol. 11, e1001640 (2013).
pubmed: 24013648
pmcid: 3754886
doi: 10.1371/journal.pbio.1001640
Ludwig, A., Nichols, B. J. & Sandin, S. Architecture of the caveolar coat complex. J. Cell Sci. 129, 3077–3083 (2016).
pubmed: 27369768
pmcid: 5004899
Stoeber, M. et al. Model for the architecture of caveolae based on a flexible, net-like assembly of Cavin1 and Caveolin discs. Proc. Natl Acad. Sci. USA 113, E8069–E8078 (2016).
pubmed: 27834731
doi: 10.1073/pnas.1616838113
pmcid: 5167190
Snead, W. T. et al. BAR scaffolds drive membrane fission by crowding disordered domains. J. Cell Biol. 218, 664–682 (2019).
pubmed: 30504247
pmcid: 6363457
doi: 10.1083/jcb.201807119
Wang, S., Zhao, Z. & Rodal, A. A. Higher-order assembly of sorting nexin 16 controls tubulation and distribution of neuronal endosomes. J. Cell Biol. 218, 2600 (2019).
pubmed: 31253649
pmcid: 6683739
doi: 10.1083/jcb.201811074
Khater, I. M., Meng, F., Wong, T. H., Nabi, I. R. & Hamarneh, G. Super resolution network analysis defines the molecular architecture of caveolae and caveolin-1 scaffolds. Sci. Rep. 8, 9009 (2018).
pubmed: 29899348
pmcid: 5998020
doi: 10.1038/s41598-018-27216-4
Liu, L. & Pilch, P. F. A critical role of cavin (polymerase I and transcript release factor) in caveolae formation and organization. J. Biol. Chem. 283, 4314–4322 (2008).
pubmed: 18056712
doi: 10.1074/jbc.M707890200
Ariotti, N. et al. Modular detection of GFP-labeled proteins for rapid screening by electron microscopy in cells and organisms. Dev. Cell 35, 513–525 (2015).
pubmed: 26585296
doi: 10.1016/j.devcel.2015.10.016
Hetmanski, J. H. R. et al. Membrane tension orchestrates rear retraction in matrix-directed cell migration. Dev. Cell 51, 460–475 (2019).
pubmed: 31607653
pmcid: 6863396
doi: 10.1016/j.devcel.2019.09.006
Pelkmans, L. & Zerial, M. Kinase-regulated quantal assemblies and kiss-and-run recycling of caveolae. Nature 436, 128 (2005).
pubmed: 16001074
doi: 10.1038/nature03866
Siahaan, V. et al. Kinetically distinct phases of tau on microtubules regulate kinesin motors and severing enzymes. Nat. Cell Biol. 21, 1086–1092 (2019).
pubmed: 31481789
doi: 10.1038/s41556-019-0374-6
Tan, R. et al. Microtubules gate tau condensation to spatially regulate microtubule functions. Nat. Cell Biol. 21, 1078–1085 (2019).
pubmed: 31481790
pmcid: 6748660
doi: 10.1038/s41556-019-0375-5
Drechsler, H., Xu, Y., Geyer, V.F., Zhang, Y. & Diez, S. Multivalent electrostatic microtubule-interactions of synthetic peptides are sufficient to mimic advanced MAP-like behaviour. Mol. Biol. Cell 30, mbcE19050247 (2019).
doi: 10.1091/mbc.E19-05-0247
Das, S., Eisen, A., Lin, Y.-H. & Chan, H. S. A lattice model of charge-pattern-dependent polyampholyte phase separation. J. Phys. Chem. B 122, 5418–5431 (2018).
pubmed: 29397728
doi: 10.1021/acs.jpcb.7b11723
Borgia, A. et al. Extreme disorder in an ultrahigh-affinity protein complex. Nature 555, 61–66 (2018).
pubmed: 29466338
pmcid: 6264893
doi: 10.1038/nature25762
Lin, Y.-H., Forman-Kay, J. D. & Chan, H. S. Sequence-specific polyampholyte phase separation in membraneless organelles. Phys. Rev. Lett. 117, 178101 (2016).
pubmed: 27824447
doi: 10.1103/PhysRevLett.117.178101
Lin, Y.-H., Forman-Kay, J. D. & Chan, H. S. Theories for sequence-dependent phase behaviors of biomolecular condensates. Biochemistry 57, 2499–2508 (2018).
pubmed: 29509422
doi: 10.1021/acs.biochem.8b00058
Feng, H., Zhou, B. R. & Bai, Y. Binding affinity and function of the extremely disordered protein complex containing human linker histone H1.0 and its chaperone ProTalpha. Biochemistry 57, 6645–6648 (2018).
pubmed: 30430826
doi: 10.1021/acs.biochem.8b01075
Pak, C. W. et al. Sequence determinants of intracellular phase separation by complex coacervation of a disordered protein. Mol. Cell 63, 72–85 (2016).
pubmed: 27392146
pmcid: 4973464
doi: 10.1016/j.molcel.2016.05.042
Zeno, W. F. et al. Synergy between intrinsically disordered domains and structured proteins amplifies membrane curvature sensing. Nat. Commun. 9, 4152 (2018).
pubmed: 30297718
pmcid: 6175956
doi: 10.1038/s41467-018-06532-3
Zeno, W. F. et al. Molecular mechanisms of membrane curvature sensing by a disordered protein. J. Am. Chem. Soc. 141, 10361–10371 (2019).
pubmed: 31180661
pmcid: 6610580
doi: 10.1021/jacs.9b03927
Busch, D. J. et al. Intrinsically disordered proteins drive membrane curvature. Nat. Commun. 6, 7875 (2015).
pubmed: 26204806
doi: 10.1038/ncomms8875
Yoo, H., Triandafillou, C. & Drummond, D. A. Cellular sensing by phase separation: using the process, not just the products. J. Biol. Chem. 294, 7151–7159 (2019).
pubmed: 30877200
pmcid: 6509497
doi: 10.1074/jbc.TM118.001191
Aoki, T., Hagiwara, H., Matsuzaki, T., Suzuki, T. & Takata, K. Internalization of caveolae and their relationship with endosomes in cultured human and mouse endothelial cells. Anat. Sci. Int. 82, 82–97 (2007).
pubmed: 17585564
doi: 10.1111/j.1447-073X.2006.00160.x
Boucrot, E., Howes, M. T., Kirchhausen, T. & Parton, R. G. Redistribution of caveolae during mitosis. J. Cell Sci. 124, 1965–1972 (2011).
pubmed: 21625007
pmcid: 3104031
doi: 10.1242/jcs.076570
Jung, W. et al. Cell-free formation and interactome analysis of caveolae. J. Cell Biol. 217, 2141–2165 (2018).
pubmed: 29716956
pmcid: 5987714
doi: 10.1083/jcb.201707004
Pelkmans, L., Burli, T., Zerial, M. & Helenius, A. Caveolin-stabilized membrane domains as multifunctional transport and sorting devices in endocytic membrane traffic. Cell 118, 767–780 (2004).
pubmed: 15369675
doi: 10.1016/j.cell.2004.09.003
Shvets, E., Bitsikas, V., Howard, G., Hansen, C. G. & Nichols, B. J. Dynamic caveolae exclude bulk membrane proteins and are required for sorting of excess glycosphingolipids. Nat. Commun. 6, 6867 (2015).
pubmed: 25897946
doi: 10.1038/ncomms7867
Murray, D. H. et al. An endosomal tether undergoes an entropic collapse to bring vesicles together. Nature 537, 107–111 (2016).
pubmed: 27556945
pmcid: 5142606
doi: 10.1038/nature19326
Miskei, M. et al. Fuzziness enables context dependence of protein interactions. FEBS Lett. 591, 2682–2695 (2017).
pubmed: 28762260
doi: 10.1002/1873-3468.12762
Olsen, J. G., Teilum, K. & Kragelund, B. B. Behaviour of intrinsically disordered proteins in protein–protein complexes with an emphasis on fuzziness. Cell Mol. Life Sci. 74, 3175–3183 (2017).
pubmed: 28597296
pmcid: 5533869
doi: 10.1007/s00018-017-2560-7
Tompa, P. & Fuxreiter, M. Fuzzy complexes: polymorphism and structural disorder in protein–protein interactions. Trends Biochem. Sci. 33, 2–8 (2008).
pubmed: 18054235
doi: 10.1016/j.tibs.2007.10.003
Richter, T. et al. High-resolution 3D quantitative analysis of caveolar ultrastructure and caveola-cytoskeleton interactions. Traffic 9, 893–909 (2008).
pubmed: 18397183
doi: 10.1111/j.1600-0854.2008.00733.x
Catanzariti, A. M., Soboleva, T. A., Jans, D. A., Board, P. G. & Baker, R. T. An efficient system for high‐level expression and easy purification of authentic recombinant proteins. Protein Sci. 13, 1331–1339 (2004).
pubmed: 15096636
pmcid: 2286746
doi: 10.1110/ps.04618904
Berrow, N. S. et al. A versatile ligation-independent cloning method suitable for high-throughput expression screening applications. Nucleic Acids Res. 35, e45–e45 (2007).
pubmed: 17317681
pmcid: 1874605
doi: 10.1093/nar/gkm047
Aboulaich, N., Vainonen, J. P., Stralfors, P. & Vener, A. V. Vectorial proteomics reveal targeting, phosphorylation and specific fragmentation of polymerase I and transcript release factor (PTRF) at the surface of caveolae in human adipocytes. Biochem. J. 383, 237–248 (2004).
pubmed: 15242332
pmcid: 1134064
doi: 10.1042/BJ20040647
Angelova, M. I. & Dimitrov, D. S. Liposome electroformation. Faraday Discuss. Chem. Soc. 81, 303–311 (1986).
doi: 10.1039/dc9868100303
Emsley, P. & Cowtan, K. Coot: model-building tools for molecular graphics. Acta Crystallogr D Biol. Crystallogr 60, 2126–2132 (2004).
pubmed: 15572765
doi: 10.1107/S0907444904019158
Adams, P. D. et al. PHENIX: a comprehensive Python-based system for macromolecular structure solution. Acta Crystallogr D Biol. Crystallogr 66, 213–221 (2010).
pubmed: 20124702
pmcid: 2815670
doi: 10.1107/S0907444909052925