Recent Publications

Paul, D., Stern, O., Vallis, Y., Dhillon, J., Buchannan, A., McMahon, H. (2023) Cell surface protein aggregation triggers endocytosis to maintain plasma membrane proteostasis. Nature Communications (article)

Henri-Fran�ois Renard, Mijo Simunovic, Jo�l Lemi�re, Emmanuel Boucrot, Maria Daniela Garcia-Castillo, Senthil Arumugam, Val�rie Chambon, Christophe Lamaze, Christian Wunder, Anne K. Kenworthy, Anne A. Schmidt, Harvey T. McMahon, C�cile Sykes, Patricia Bassereau & Ludger Johannes (2015) Endophilin-A2 functions in membrane scission in clathrin-independent endocytosis. Nature. 2015 Jan 22;517(7535):493-6. doi: 10.1038/nature14064. Epub 2014 Dec 17. PMID: 25517096 (abstract) (pdf including supplementary figures)
This Nature letter extends the observations of the article below, providing evidence that bacterial toxins, shiga and cholera, take advantage of an endophilin-dependent endocytic pathway to enter cells. The paper also finds that endophilin-A2 acts together with dynamin and actin.

Emmanuel Boucrot, Antonio P. A. Ferreira, Leonardo Almeida-Souza, Sylvain Debard, Yvonne Vallis, Gillian Howard, Laetitia Bertot, Nathalie Sauvonnet & Harvey T. McMahon (2015) Endophilin marks and controls a clathrin-independent endocytic pathway. Nature. 2015 Jan 22;517(7535):460-5. doi: 10.1038/nature14067. Epub 2014 Dec 17. PMID: 25517094 (abstract) (pdf including supplementary figures) (Video1) (Video2) (Video3) (Video4) (Video5)

This Nature article shows that endophilin-dependent endocytosis is a novel endocytic pathway distinct from clathrin-mediated endocytosis. This pathway is triggered by binding of ligands to cargo receptors, and requires the proteins dynamin and actin. It occurs primarily from the leading edges of cells where lamellipodin and the lipid PtdIns(3,4)P2 ensures endophilin enrichment. This form of endocytosis is shown to mediate the uptake of several physiological and disease-relevant receptors including G-protein-coupled receptors and receptor tyrosine kinases. See also BAR Superfamily Home Page

Safa Lucken-Ardjomande H�sler, Yvonne Vallis, Helen E. Jolin, Andrew N. McKenzie and Harvey T. McMahon (2014) GRAF1a is a brain-specific protein that promotes lipid droplet clustering and growth, and is enriched at lipid droplet junctions. J. Cell Sci. 2014 Nov 1;127(21):4602-19. doi: 10.1242/jcs.147694. Epub 2014 Sep 4. PMID: 25189622; PMCID: PMC4215711 (abstract) (pdf including supplementary figures) (Video1, large version) (Video2, large version) (Video3, large version) (Video4, large version)

Lipid droplets are found in all cell types. Normally present at low levels in the brain, they accumulate in tumours and are associated with neurodegenerative diseases. However, little is known about the mechanisms controlling their homeostasis in the brain. We found that GRAF1a, the longest GRAF1 isoform (GRAF1 is also known as ARHGAP26), was enriched in the brains of neonates. Endogenous GRAF1a was found on lipid droplets in oleic-acid-fed primary glial cells. Exclusive localization required a GRAF1a-specific hydrophobic segment and two membrane-binding regions, a BAR and a PH domain. Overexpression of GRAF1a promoted lipid droplet clustering, inhibited droplet mobility and severely perturbed lipolysis following the chase of cells overloaded with fatty acids. Under these conditions, GRAF1a concentrated at the interface between lipid droplets (shown as green spots on the blue rings in picture on the left). Although GRAF1-knockout mice did not show any gross abnormal phenotype, the total lipid droplet volume that accumulated in GRAF1-/- primary glia upon incubation with fatty acids was reduced compared to GRAF1+/+ cells. These results provide additional insights into the mechanisms contributing to lipid droplet growth in non-adipocyte cells, and suggest that proteins with membrane sculpting BAR domains play a role in droplet homeostasis. See also BAR Superfamily Home Page

Michael M. Kozlov, Felix Campelo, Nicole Liska, Leonid V. Chernomordik, Siewert J. Marrink and Harvey T. McMahon (2014) Mechanisms shaping cell membranes. Current Opinions in Cell Biology 29, 53-60. PMID: 24747171 (abstract) (pdf)

This is a cross between a paper and a review. It is essentially a reply to hypothesis by the Fletcher lab that membrane curvature is driven by protein crowding and not by insertions. It includes new data from Nicole Liska on curvature driven by crowding, new calculations on the effectiveness of this method by Kozlov and Campelo and new dynamic simulation of membranes by Marrink. Stachowiak et al 2012 claimed that protein crowding was the primary mechanism by which epsin deforms liposomes. Here we show that crowding can only make a contribution to bending but membrane insertion of the amphipathic helix is the primary factor. The picture on the left shows the membrane interaction surface of epsin1 ENTH domain viewed from the membrane. The amphipathic helix, which inserts into the membrane is shown in dark green with amino acid side chains included. Space filled representation of the footprint with side chains included for the whole protein, overlaid with a carbon atom lattice (to facilitate area calculations), showing that the amphipathic helix occupies approximately 1/3rd of the total projection area. See also BAR Superfamily Home Page

Tetsuya Takeda, Iain M. Robinson, Matthew M. Savoian, John R. Griffiths, Anthony D. Whetton, Harvey T. McMahon and David M. Glover (2013) Drosophila F-BAR protein Syndapin contributes to coupling the plasma membrane and contractile ring in cytokinesis. Open Biol. 2013 Aug 7;3(8):130081. doi: 10.1098/rsob.130081 PMID: 23926047, PMC3758542 (abstract) (pdf including supplementary figures)

This paper reports for the first time that an F-BAR protein Syndapin (also called Pacsin) functions at the interface between membrane and the actomyosin contractile ring during cytokinesis of animal cells. See also BAR Superfamily Home Page

Michael Meinecke, Emmanuel Boucrot, Gamze Camdere, Wai-Ching Hon, Rohit Mittal, and Harvey T McMahon (2013) Cooperative recruitment of Dynamin and BAR domain-containing proteins leads to GTP-dependent membrane scission. J. Biol. Chem. 288, 6651-6661 PMID:23297414, PMC3585104 (abstract) (pdf including supplementary figures)

This paper reports the cooperativity between dynamin and N-BAR domain-containing proteins (amphiphysin and endophilin) for membrane binding and vesicle scission. In cells we observed that 1. N-BAR protein overexpression leads to increased dynamin recruitment 2. RNAi against N-BAR-proteins leads to a reduction in dynamin recruitment. 3. We show that this RNAi of N-BAR proteins, directly leads to an inhibition of clathrin-coated vesicle (CCV) maturation. 4. The consequence of amphiphysin RNAi on CCV formation was more dramatic than that of endophilin RNAi. The clathrin-amphiphysin interaction inhibitor, pitstop, recapitulated the phenotype, showing that this drug ultimately is an inhibitor of CCV formation because it indirectly inhibits the recruitment of dynamin. To understand the molecular basis of the observed cooperativity in vivo we developed an assay for membrane binding and scission using purified proteins and giant unilamellar vesicles (GUVs). In vitro we observed that 1. Binding of either endophilin or amphiphysin to PIP2-containing membranes (GUVs) was enhanced when dynamin (or simply the dynamin- PRD) was bound to their SH3 domains. 2. In a similar manner dynamin interaction with membranes was stronger when its PRD binds the SH3 domain of either endophilin or amphiphysin. 3. This N-BAR dependence for dynamin recruitment to membranes for the first time recapitulates in vivo observations of adaptor dependence for dynamin recruitment to sites of endocytosis. We also recapitulate dynamin mediated vesicle scission in the presence of GTP and show this is enhanced in the presence of endophilin or amphiphysin. This new assay for vesicle scission is based on the reduction in the size of GUVs with time in the presence of GTP, indicating vesiculation. We report a number of surprises: 1. RNAi against endophilin has no effect on transferrin endocytosis in BSC1 cells (Fig1E), which is contrary to implications in the literature, and implies that endophilin may have little to do with clathrin-coated vesicle maturation in cells. 2. Endophilin overexpression leads to increased dynamin membrane recruitment, while dynamin RNAi leads to reduced endophilin membrane recruitment. While these results reinforce the evidence for cooperativity between the molecules we also observed that endophilin RNAi appeared to lead to reduced dynamin expression (an unexpected result). 3. It has not been previously observed that effectors are needed to either recruit dynamin or to enhance dynamin activity in vitro. Our observations imply that GUV membranes more closely resemble the cellular environment in which these cooperative effects are observed. See BAR Superfamily Home Page for details of endophilin and amphiphysin See also Dynamin web pages.

Katia Cortese, Mark T. Howes, Richard Lundmark, Erica Tagliatti, Paola Bagnato, Annalisa Petrelli, Maria Bono, Harvey T. McMahon, Robert G. Parton and Carlo Tacchetti (2013) The HSP90 inhibitor geldanamycin perturbs endosomal structure and drives recycling ErbB2 and transferrin to modified MVBs/lysosomal compartments. Mol Biol Cell. 2013 Jan;24(2):129-44. doi: 10.1091/mbc.E12-04-0282. Epub 2012 Nov 14. PMID: 23154999, PMC3541960 (abstract) (pdf) (Movie1) (Movie2) (Movie3) (Movie4) (Movie5) A study on how the anti-cancer drug, geldanamycin, affects endocytic trafficking

Maria Jose Sanchez-Barrena, Yvonne Vallis, Menna R. Clatworthy, Gary J. Doherty, Dmitry B. Veprintsev, Philip R. Evans and Harvey T. McMahon (2012) Bin2 Is a Membrane Sculpting N-BAR Protein That Influences Leucocyte Podosomes, Motility and Phagocytosis. PLoS One. 2012;7(12):e52401. doi: 10.1371/journal.pone.0052401. Epub 2012 Dec 20. PMID: 23285027 PMCID: PMC3527510 (abstract) (pdf including supplementary figures) (Movie 1) (Movie2)

This paper reports the structure and functional analysis of Bin2/BRAP BAR domain-containing protein Bin2 is only expressed in leucocytes and we have found it to be implicated in cell motility, adhesion and phagocytosis that are controlled by actin and membrane remodelling processes. We have solved the structure of Bin2 N-BAR domain and we have shown that this domain enables the protein to tubulate membranes in vivo and in vitro. Compared to previously characterized N-BAR-containing proteins, Bin2 lacks an SH3 domain to bind dynamin and in contrast, it contains a poly-proline region which we show interacts with endophilin, and PIX. We have found that Bin2 is a new component of the podosomal machinery. Bin2 N-BAR is responsible for its targeting to these actin rich structures and its C-terminus permits Bin2 to interact with SH3 domain containing proteins such as alpha-PIX and Endophilin A2. Manipulation of Bin2 protein levels affects podosome formation as it has been shown for PIX and its binding partner PAK. Furthermore, Bin2 is at the leading edge of migrating leukocytes and favours cell movement. Additionally, Bin2 is also found at the phagocytic cup of macrophages and inhibits phagocytosis. In summary, Bin2 is a N-BAR protein enriched in leucocytes and implicated in actin rearrangements associated with leucocyte morphogenesis. See BAR Superfamily Home Page for details of endophilin and amphiphysin

Emmanuel Boucrot, Adi Pick, Gamze Camdere, Nicole Liska, Emma Evergren, Harvey T. McMahon, Michael M. Kozlov (2012) Membrane fission is promoted by insertion of amphipathic helices and is restricted by crescent BAR domains. Cell Mar 30;149(1):124-36. PMID: 22464325 (abstract) (pdf including supplementary materials and figures)

Violeta Chitu, Viorel Nacu, Julia F. Charles, William M. Henne, Harvey T. McMahon, Sayan Nandi, Halley Ketchum, Renee Harris, Mary C. Nakamura and E. Richard Stanley (2012) PSTPIP2 deficiency in mice causes osteopenia and increased differentiation of multipotent myeloid precursors into osteoclasts. Blood 120, 3126-3135. [Epub ahead of print Aug24] PMID: 22923495 (abstract) (pdf including supplementary figures)

Chun-Liang Lai, Christine C. Jao, Edward Lyman, Jennifer L. Gallop, Brian J. Peter, Harvey T. McMahon, Ralf Langen and Gregory A. Voth (2012) Membrane Binding and Self-Association of the Epsin N-Terminal Homology Domain. Journal of Molecular Biology 423, 800-817. PMID: 22922484 (abstract) (pdf)

Bjorn Moren, Claudio Shah, Mark T. Howes, Nicole L. Schieber, Harvey T. McMahon, Robert G. Parton, Oliver Daumke and Richard Lundmark (2012) EHD2 regulates caveolar dynamics via ATP-driven targeting and oligomerization. Mol. Biol. Cell Apr;23(7):1316-29. PMID: 22323287, PMCID: PMC3315815 (abstract) (pdf including supplementary materials and figures) (smaller versions of movies S1 635KB, S2 258KB, S3A 5MB, S3B 3.6MB, S4A 6.2MB, S4B 3.8MB)(full size movies S1 2.4MB, S2 1.2MB, S3A 24.3MB, S3B 15.2MB, S4A 35.7MB, S4B 30.3MB)

Gary J. Doherty, Monika K. Ahlund, Mark T. Howes, Bjorn Moren, Robert G. Parton, Harvey T. McMahon and Richard Lundmark (2011) The endocytic protein GRAF1 is directed to cell-matrix adhesion sites and regulates cell spreading. Mol. Biol. Cell. Nov;22(22):4380-4389. PMID: 21965292 (abstract) (pdf including supplementary materials and figures) (supplementary Movies 1, 2, 3, 4, 5)

Artur Llobet, Jennifer L. Gallop, Jemima E. Burden, Gamze Camdere, Priya Chandra, Yvonne Vallis, Collin R. Hopkins, Leon Lagnado, Harvey T. McMahon (2011) Endophilin drives the fast mode of vesicle retrieval in a ribbon synapse. J. Neurosci. Jun 8;31(23):8512-9. PMID: 21653855(abstract) (pdf including supplementary materials and figures)

Jean-Philippe Richard, Evgenia Leikina, Ralf Langen, William Mike Henne, Margarita Popova, Tamas Balla, Harvey T McMahon, Michael Kozlov and Leonid Chernomordik (2011) Intracellular curvature generating proteins in cell-to-cell fusion. Biochemical J. Dec 1;440(2):185-193. Epub 2011 Sept 7. PMID: 21895608 PMCID: PMC3216009 (abstract) (pdf)

Sascha Martens, Harvey T. McMahon (2011) C2 domains and membrane fusion. Curr. Top. Membr. 68:141-59. PMID: 21771498 (pdf)

Harvey T. McMahon, Emmanuel Boucrot (2011) Molecular mechanism and physiological functions of clathrin-mediated endocytosis. Nat. Rev. Mol. Cell Biol. Jul 22;12(8):517-33. doi: 10.1038/nrm3151. PMID: 21779028 (abstract) (pdf)

Emmanuel Boucrot, Harvey T. McMahon (2011) [Nucleation of clathrin-coated pits - 'membrane sculptors' at work]. Med. Sci. (Paris). Feb;27(2):122-5. Epub 2011 Mar 8. French. PMID: 21382316 (abstract) (pdf)

Misha M. Kozlov, Harvey T. McMahon and Leonid V. Chernomordik (2010) Protein-driven membrane stresses in fusion and fission. Trends Biochem. Sci. 35, 699-706. [Epub ahead of print Jul 15 2010] PubMed PMID: 20638285. (abstract) (pdf)

William Mike Henne*, Emmanuel Boucrot*^, Michael Meinecke, Emma Evergren,Yvonne Vallis, Rohit Mittal and Harvey T. McMahon^ (2010) FCHo Proteins are Nucleators of Clathrin-Mediated Endocytosis. Science 328, 1281-1284. (*joint first authors,^joint corresponding authors) (Published online 6 May 2010; 10.1126/science.1188462) (abstract) (pdf including supplementary materials and figures) (movie1, increased CCV budding with FCHo2 overexpression) (movie2, simulation of membrane bending by FCHo proteins) PMID: 20448150; PMCID: PMC2883440 (Faculty of 1000 Exceptional rating) (News and Views in Nature) (Science Signalling, editors' choice)(Nature Reviews in Molecular Cell Biology, Research Highlight)

This is first membrane curvature effector protein to define sites where clathrin-coated vesicles will bud. It also appears to be the first 'coat-accessory protein' recruited recruited to the plasma membrane for the initiation of CCV formation here. Its expression levels correlate with the number of budding events and thus ligand uptake. The proteins are ubiquitously expressed and are shown to function in fibroblasts and in synapses. See F-BAR homepage for structure details. See endocytosis.org homepage for research on other endocytic proteins.

Misha M. Kozlov, Harvey T. McMahon and Leonid V. Chernomordik (2010) Protein-driven membrane stresses in fusion and fission. Trends Biochem Sci. 35, 699-706. [Epub ahead of print Jul 15 2010] PubMed PMID: 20638285. (abstract) (pdf)

Mark T. Howes, Matthew Kirkham, James Riches, Katia Cortese, Piers J. Walser, Fiona Simpson, Michelle M. Hill, Alun Jones, Richard Lundmark, Margaret R. Lindsay, Delia J. Hernandez-Deviez, Gordana Hadzic, Adam McCluskey, Rumasia Bashir, Libin Liu, Paul Pilch, Harvey McMahon, Phillip J. Robinson, John F. Hancock, Satyajit Mayor and Robert G. Parton (2010) Clathrin-independent carriers form a high capacity endocytic sorting system at the leading edge of migrating cells. J. Cell Biol. Aug 23;190(4):675-91. Epub 2010 Aug 16. PMID: 20713605; PMCID: PMC2928008. (abstract) (pdf including supplemental figures)

Ji-Young Youn, Helena Friesen, Takuma Kishimoto, William Mike Henne, Christoph F. Kurat, Wei Ye, Derek F. Ceccarelli, Frank Sicheri, Sepp D. Kohlwein, Harvey T. McMahon, Brenda J. Andrews (2010) Dissecting BAR domain function in the yeast Amphiphysins Rvs161 and Rvs167 during endocytosis. Mol. Biol. Cell. Sep;21(17):3054-3069. [Epub ahead of print Jul 7 2010] PMID: 20610658; PubMed Central PMCID: PMC2929998 (abstract) (pdf including supplemental material and figures)

Harvey T. McMahon, Misha Kozlov and Sascha Martens (2010) Membrane Curvature in Synaptic Vesicle Fusion and Beyond. Cell 140, 601-605. DOI 10.1016/j.cell.2010.02.017 PMID 20211126 (abstract) (pdf)

This essay presents the case why: A. high transient membrane curvature is intrinsically fusogenic B. synaptotagmin should be considered as a core fusion protein along with the SNARE proteins C. all membrane fusion events likely involve a high curvature intermediate D. many of the components in SNARE mediated fusion will contribute to curvature, including the SNARE proteins E. a point source of fusion is created by high curvature and should lead to non-leaky membrane fusion. See Synaptotagmin homepage for how synaptotagmin works

Rohit Mittal, Sew-Yeu Peak-Chew, Robert S Sade, Yvonne Vallis and Harvey T McMahon (2010) The acetyltransferase activity of the bacterial toxin YopJ of Yersinia is activated by eukaryotic host cell inositol hexakisphosphate. J. Biol. Chem. 285, 19927-199234. [Epub ahead of print 29Apr 2010] DOI 10.1074/jbc.M110.126581 PMID: 20430892 (abstract) (pdf including supplemental material and figures)

The bacterial toxin YopJ from Yersinia acetylates serine and threonine residues on important molecules in the MAP kinase and NFkappaB signalling pathways (see Mittal et al 2006, below). In this paper we now establish that YopJ from Yersinia and AvrA from Salmonella are activated by inositol hexakisphosphate (IP6) present in the eukaryotic target cell. Our results suggest that YopJ-family enzymes are quiescent in the bacterium where they are synthesized because bacteria lack IP6; once injected into mammalian cells by the pathogen these toxins bind host cell IP6, are activated and deregulate the MAPK and NFkappaB signaling pathways thereby subverting innate immunity. See Black Plague Home Page that gives the background and further information on this paper.

Christine C. Jao, Balachandra G. Hegde, Jennifer L. Gallop, Prabhavati B. Hegde, Harvey T. McMahon, Ian S. Haworth and Ralf Langen (2010) Roles of amphipathic helices and the BAR domain of endophilin in membrane curvature generation. J. Biol. Chem. 285, 20164-70. [Epub ahead of print Apr 23, 2010] PMID: 20418375 (pdf)

(Alexander J. Groffen^, Sascha Martens^), Roc�o D�ez Arazola, L. Niels Cornelisse, Natalia Lozovaya, Arthur P. H. de Jong, Natalia A. Goriounova, Ron L. P. Habets, Yoshimi Takai, J. Gerard Borst, Nils Brose, (Harvey T. McMahon*, and Matthijs Verhage*) (2010) Doc2b Is a High-Affinity Ca2+ Sensor for Spontaneous Neurotransmitter Release. Science (article), 327, 1614-1618[First published online 11 February 2010 in Science Express Research Articles] [DOI: 10.1126/science.1183765] ^joint first and corresponding authors and *joint senior authors. PMID: 20150444 (pdf including supplementary material and figures) (CNCR news) (Faculty of 1000 Exceptional rating)
           Here we have found a calcium sensor for spontaneous neurotransmitter release in the brain. It has a higher calcium affinity than synaptotagmin1 and thus can respond to much lower calcium rises. In vitro it promotes SNARE-dependent membrane fusion by promoting extreme local membrane curvature. This sensor competes with synaptotagmin, allowing us to predict that in vivo the properties of an individual vesicle fusion event will be a product of the different calcium-sensors present at the vesicle release site.

Felix Campelo, Gur Fabrikant, Harvey T. McMahon and Michael M. Kozlov (2009) Modeling membrane shaping by proteins: Focus on EHD2 and N-BAR domains FEBS Lett. 584, 1830-1839. [Epub ahead of print 16Oct 2009] PMID: 19836393 (abstract) (pdf)

Ahead of the curve A Nature News and Views feature that highlights our work (16 July issue Nature 2009) (pdf)

Gary J. Doherty and Harvey T. McMahon (2009) Mechanisms of Endocytosis. Annual Review in Biochemistry. 78, 857-902. [First published online as a Review in Advance on March 24, 2009] PMID: 19317650 (abstract) (pdf)

Rohit Mittal and Harvey T. McMahon (2009) Arrestins as adaptors for ubiquitination in endocytosis and sorting. EMBO Rep. 10, 41-43. [Epub ahead of print Dec 2008] PMID: 19057574 (abstract) (pdf)

Richard Lundmark, Gary J. Doherty, Mark T. Howes, Katia Cortese, Yvonne Vallis, Robert G. Parton and Harvey T. McMahon (2008) The GTPase-activating protein GRAF1 regulates the CLIC/GEEC endocytic pathway. Current Biology 18, 1802-1808. PMID: 19036340; PMC2726289; doi: 10.1016/j.cub.2008.10.044. (abstract) (pdf including supplementary figures)
           A clathrin-independent tubulovesicular endocytic pathway dependent on the BAR-domain protein GRAF1.

Eberth A, Lundmark R, Gremer L, Dvorsky R, Koessmeier KT, McMahon HT, and Ahmadian MR. (2009) A BAR domain-mediated autoinhibitory mechanism for RhoGAPs of the GRAF family. Biochem. J. 417, 371-377. PMID: 18954304 (abstract) (pdf including supplementary figures)

Kenneth L. Madsen, Jacob Eriksen, Laura Milan-Lobo, Daniel S. Han, Masha Y. Niv, Ina Ammendrup-Johnsen, Ulla Henriksen, Vikram K. Bhatia, Dimitrios Stamou, Harald H. Sitte, Harvey T. McMahon, Harel Weinstein and Ulrik Gether (2008) Membrane Localization is Critical for Activation of the PICK1 BAR Domain. Traffic 9, 1327�1343. PMID: 18466293 (abstract) (pdf including supplementary material)

Kara L. Lynch, Roy R.L. Gerona, Dana M. Kielar, Sascha Martens, Harvey T. McMahon and Thomas F.J. Martin (2008) Synaptotagmin-1 utilizes membrane bending and SNARE binding to drive fusion pore expansion. Molecular Biology of the Cell 19, 5093-5103. [Epub ahead of print Sept17 2008] PMID: 18799625 (abstract) (pdf including supplementary material) (Faculty of 1000 Biology Reviewed)

Richard Lundmark, Gary J. Doherty, Yvonne Vallis, Brian J. Peter and Harvey T. McMahon (2008) Arf family GTP loading is activated by, and generates, positive membrane curvature. Biochemical J. 414, 189-194. doi:10.1042/BJ20081237 PMID: 18597672 (abstract) (pdf) (commentary by Julia Donaldson, NIH)

Felix Campelo, Harvey T. McMahon and Misha M. Kozlov (2008) The hydrophobic insertion mechanism of membrane curvature generation by proteins. Biophys J. 95, 2325-2339. [Epub ahead of print May 30] doi:10.1529/biophysj.108.133173, PMID: 18515373 (abstract) (pdf)

Sascha Martens and Harvey T. McMahon (2008) Mechanisms of membrane fusion: disparate players and common principles. Nature Reviews Molecular Cell Biology, 9, 543-556 (July issue). Advance online publication, 21 May 2008 | doi:10.1038/nrm2417, PMID: 18496517 (abstract) (pdf)

Gary J. Doherty and Harvey T. McMahon (2008) Mediation, Modulation, and Consequences of Membrane-Cytoskeleton Interactions. Annual Review in Biophysics 37, 65-95. PMID: 18573073 First published online as a Review in Advance on December 3, 2007 (doi:10.1146/annurev.biophys.37.032807.125912) (pdf) (Ann Rev reprint)

Lene E. Olesen, Eva M. Schmid, Marijn G. J. Ford, Yvonne Vallis, M. Madan Babu, Peter Li, Ian G. Mills, Philip R. Evans, (Harvey T. McMahon and Gerrit J.K. Praefcke) (2008) Solitary and repetitive binding motifs for the AP2 complex alpha-appendage in amphiphysin and other accessory proteins. J. Biol. Chem. 283, 5099-5109. PMID: 17986441 First published on line in 2007, JBC Papers in Press, November 6, 2007, DOI 10.1074/jbc.M708621200 (abstract) (pdf and supplementary material)

Oliver Daumke, Richard Lundmark, Yvonne Vallis, Sascha Martens, P. Jonathan G. Butler and Harvey T. McMahon (2007) Architectural and mechanistic insights into an EHD ATPase involved in membrane remodelling. Nature, 449 923-927. PMID: 17914359 (doi:10.1038/nature06173, online 3 Oct issue). (pdf) (abstract) (Supplementary Figures) (coordinates of proposed EHD oligomer, made from 4 dimers) (Faculty of 1000 - evaluation 6.5)

Eva M. Schmid and Harvey T. McMahon (2007) Integrating molecular and network biology to decode endocytosis. Nature, 448, 883-888. (doi:10.1038/nature06031, 23Aug issue). PMID: 17713526 (pdf) (abstract) (Supplementary Material)

William Mike Henne, Helen M. Kent, Marijn G.J. Ford, Balachandra G. Hegde, Oliver Daumke, P. Jonathan G. Butler, Rohit Mittal, Ralf Langen, Philip R. Evans, and Harvey T. McMahon (2007) Structure and Analysis of FCHo2 F-BAR Domain: a Dimerising and Membrane Recruitment Module that Effects Membrane Curvature. Structure 15, 839-852. PMID: 17540576 doi:10.1016/j.str.2007.05.002 (pdf) (Supplementary material) (MRC highlight)

Sascha Martens, Michael M. Kozlov and Harvey T. McMahon (2007) How Synaptotagmin Promotes Membrane Fusion. Science. Published online 3 May 2007; 10.1126/science.1142614
Print Version: Science 2007, 316, 1205-1208 (25May) PMID: 17478680 (pdf including Supplemental material) (abstract from Science Website) (reprint from Science Website) (Highlight in Nature Structure and Molecular Biology: Syts throw a curve, 14, 467 (2007) doi:10.1038/nsmb0607-467 (pdf)) (Highlight in October 15 issue 2007 Journal of Physiology)
            This paper shows that synaptotagmin-1, the major calcium sensor for synaptic vesicle exocytosis, induces extreme positive membrane curvature upon calcium-induced binding to membranes. It does this by inserting hydrophobic residues into the hydrophobic phase of the membrane. It has previously been shown that synaptotagmin-1 has to bind to membranes in order to promote synaptic vesicle exocytosis but the downstream effect of this membrane binding was unknown. We were thus the first to assign a functional outcome to calcium-induced membrane-binding by synaptotagmin and show that it is the extreme curvature induced by this protein that leads to membrane fusion. We went on to show that other synaptotagmin-related molecules also induce membrane curvature and showed that the calcium-dependent induction of membrane curvature by synaptotagmin-1 is essential for the promotion of SNARE-dependent membrane fusion. According to our model the extreme membrane curvature induced by synaptotagmin-1 results in the induction of curvature stress thereby reducing the high energy barrier for membrane fusion. Synaptotagmin-related molecules with multiple linked C2 domains are ubiquitous and our results suggest that the promotion of membrane fusion by multiple C2 domain containing proteins by the induction extreme positive curvature is a widespread phenomenon in biology. See Synaptotagmin Home Page

Anne Burtey, Eva M. Schmid, Marijn G.J. Ford, Joshua Z. Rappoport, Mark G.H. Scott, Stefano Marullo, Sanford M. Simon, Harvey T. McMahon and Alexandre Benmerah (2007) The conserved IVF motif couples activation state and endocytic functions of beta-arrestins. Traffic 8, 914-931(Epub 2007 Jun 4) PMID: 17540576 (pdf) (abstract)
            Beta-arrestins play a central role in the regulation of G protein coupled receptors (GPCRs) and in their targeting for endocytosis. We previously solved the structure of the C-terminus of beta2-arrestin with beta-adaptin (Schmid et al 2006, see below). The adaptin interaction drives the recruitment of beta-arrestins-GPCR complexes into clathrin-coated pits. This paper describes in vitro and in vivo studies on arrestin dynamics.

Mittal, R., Peak-Chew, S-Y. and McMahon, H.T. (2006) Acetylation of MEK2 and IkB kinase (IKK) activation loop residues by YopJ inhibits signalling. PNAS 103, 18574-18579. PMID: 17116858 (abstract) (pdf) (Sup Figs 4-7) (Faculty of 1000 -exceptional factor 9).
            This paper shows that the Yersinia toxin YopJ blocks the MAP kinase pathway and the NF-kB pathway by acetylation of serine residues in MEKs and IKKs respectively. Previously acetylation has been shown for lysine residues but here we show acetylation directly by the Yersinia toxin on serine and threonine residues in the activation loops of these kinases. These results raise the possibility that this type of acetylation may occur under non-pathogenic conditions and may regulate protein function in eukaryotic cells.

Schmid, E.M., Ford, M.G.J., Burtey, A., Praefcke, G.J.K., Peak-Chew, S-Y., Mills, I.G., Benmerah, A. and McMahon, H.T. (2006) Role of the AP2 beta-appendage hub in recruiting partners for clathrin-coated vesicle assembly. PLoS Biology 4(9), e262. PMID: 16903783 DOI: 10.1371/journal.pbio.0040262 (pdf) (sup.pdf)
            This paper demonstrates the power of a multidisciplinary approach to understand the process of clathrin-mediated endocytosis. The investigation at first focuses on a comparison of the AP2 appendage interactors. By using a mass spectrometry-based proteomic approach we conclude that the “second ear” (the beta-appendage) helps widen the interaction repertoire of the AP2 complex and increases the avidity of some interaction partners. X-ray crystallography allowed us to identify two binding sites on the b-appendage, one on the top and the other on the side of the appendage, and from the structures we can see the essential interacting residues. Thus we can identify potential appendage binding motifs in other ligands. We also found a new mode of appendage interaction, where the top site interacting peptide from beta-arrestin undergoes a conformational change into a helix upon binding to the beta-appendage. This is again used by other ligands and the combination of sequence motif and structural modification gives an extra level of specificity to the interaction.

Painting by an art student, Alex McCuish from Hills Road Sixth Form College, Cambridge UK, of the protein interaction network underlying nerve function. See Interactome Home Page

This paper gives us insights into the overall design principles behind clathrin-mediated endocytosis.
Firstly, we can give a reasoning why two appendages instead of one are needed: proteins that bind to both appendages have a higher chance of being recruited. Once recruited they have reduced off-rates and therefore are more likely to stay at the site of action where they are needed and will only be replaced by higher affinity binders. Such higher affinity binders can be proteins which have more than one interaction motif for the appendages and can therefore crosslink AP2’s on the membrane via binding to multiple appendages from different AP2 complexes.
Secondly: in the progression of endocytosis there is a maturation of hubs and while ligands may initially be recruited to sites of endocytosis by interactions with AP2 appendages, yet, to be incorporated in clathrin-coated vesicles they must need to change partners and become attached to clathrin coat.
Thirdly: to maintain reversibility and dynamic instability in the process we should consider the build up of protein interactions as a network where a combination of the reaching a critical concentration (through clustering) and the energy input step defines the point of no return. We therefore see the AP2 complex and clathrin as the major control centres of clathrin-mediated endocytosis, the hubs in a dynamic pathway-network.
Finally: we propose a new type of protein interaction that cannot be described by affinities or avidities because it is a semi-solid state interaction, and to describe this we coin the term, ‘Matricity’. This describes the interaction of a clathrin coat with its membrane attachment proteins and cargo adaptors.

Gallop, J.L., Jao, C.C., Kent, H.M., Butler, P.J.G., Evans, P.R., Langen, R. and McMahon, H.T. (2006) Mechanism of endophilin N-BAR domain-mediated membrane curvature. EMBO J. 25, 2898-2910. PMID: 16763559 (abstract) (pdf) (Sup Figures) (Sup Methods)
            The crystal structure of endophilin BAR domain and electron spin resonance analysis of the N-terminal amphipathic helix in the presence of membranes. This paper shows how the N-BAR domain (amphipathic helix plus BAR domain) works as a functional unit to promote dimerisation and membrane curvature generation. An insert in the middle of the BAR domain, defines an endophilin/nadrin subfamily of BAR domains, where part of this insert forms another short amphipathic helix and also contributes to specificity of binding. See Endophilin Home Page

McMahon, H.T. and Gallop J.L. (2005) Membrane curvature and mechanisms of dynamic cell remodelling. Nature 438, 590-596. (invited review) (pdf)
            Exploratory review on the role of proteins in dynamically changing membrane conformation and the downstream effects of these changes. We address different mechanisms of curvature generation and how these interact to orchestrate membrane trafficking. See BAR Superfamily Home Page

Higgins, M and McMahon, H.T. (2005) In vitro reconstitution of discrete stages of dynamin-dependent endocytosis. Methods Enzymology 404, 597-611. (pdf)

Gallop, J.L., Butler, P.J.G. and McMahon H.T. (2005) Endophilin and CtBP/BARS are not acyl transferases in endocytosis or Golgi fission. Nature 438, 675-678. (pdf) (supplementary figures)
         Both endophilins and CtBP/BARS have been proposed to have lysophosphatidic acid acyl transferase (LPAAT) activities that can make phosphatidic acid in membranes. For endophilins this activity is thought to change the bilayer asymmetry in such a way that negative membrane curvature at the neck of a budding vesicle will be stabilised. Here we show that the LPAAT activities are contaminants associating with these proteins during purification. This now prompts a re-evaluation of the functions of these proteins. See Endophilin Home Page

Jochusch, W.J., Praefcke, G.J.K., (McMahon, H.T. and Lagnado, L.) (2005) Clathrin-dependent and clathrin-independent retrieval of synaptic vesicles in retinal bipolar cells. Neuron 46, 869-878. (abstract) (pdf).

Endocytosis in central neurons can frequently be separated into two kinetic components. Here we show that the fast component in retinal bipolar cells (time constant of 1s) is clathrin-independent and dynamin dependent. The slow component (time constant of 10s) is AP2 adaptor, clathrin and dynamin-dependent and thus is the canonical clathrin-mediated endocytic pathway.

Gallop, J.L. and McMahon H.T. (2005) BAR domains and membrane curvature: bringing your curves to the BAR. Biochem. Soc. Symp. 72, 223-231. (pdf)
Do BAR domains create membrane subdomains and how can BAR domains influence membrane trafficking? This review covers the significance of, and ways, in which BAR domains function in various contexts.

Carlton, J.G., Bujny, M.V., Peter, B.J., Oorschot, V.M.J., Rutherford, A., Arkell, R.S., Klumperman, J., McMahon, H.T. and Cullen, P.J. (2005) Sorting nexin-2 is associated with tubular elements of the early endosome, but is not essential for retromermediated endosome-to-TGN transport. J. Cell Sci. 118, 4527-4539. (pdf)
            A continuation of the collaboration with Pete Cullen, Bristol, on the role of sorting-nexins in membrane trafficking. The BAR domain of sorting-nexin2 does not tubulate membranes but is sensitive to curvature and in vivo sorting-nexin2 localises to tubular elements of endosomes.

Praefcke, G.J.K., Ford, M.G.J., Schmid, E.M., Olesen, L.E., Gallop, J.L., Peak-Chew, S-Y., Vallis, Y., Madan Babu, M., Mills, I.G. and McMahon, H.T. (2004) Evolving nature of the AP2 alpha-appendage hub during clathrin-coated vesicle endocytosis. EMBO J. 23, 4371-4383. PMID: 15496985 (abstract) (pdf) (movie of AP2 alpha-appendage with synaptojanin peptides) (supplementary material) (see also cover of same issue) (Highlight in Science 2004, v306, p1103, pdf)

This is the first of a series of network papers on trying to understand the making of a clathrin-coated vesicle as a 'series of many parallel processes'. In protein interaction networks, hubs are proteins that have disproportionately large numbers of interaction partners. For these hubs to function in a biological process it would seem that there must be sequential or spatial ordering of the interactions. In this paper, where we treat clathrin-mediated endocytosis as a module of a network, we uncover how the AP2 alpha-appendage works as an interaction hub after being concentrated at sites of endocytosis. These concentrated appendages provide a multivalent binding platform (hub) for interaction partners. Thus the partners are represented according to their relative affinities and concentrations. In this paper we also discuss the biological basis for the temporal nature of this hub and for the directionality imposed by the system. This has implications for all biological networks where similar principles are likely to underlie their functions. (AP2 alpha-appendage network hub used on journal cover Large version of EMBO J. Cover and Cover text) See also adaptor appendage domain web pages

Agnes Saint-Pol, Belen Yelamos, Mohamed Amessou, Ian G. Mills, Marc Dugast, Daniele Tenza, Peter Schu, Claude Antony, Harvey T. McMahon, Christophe Lamaze and Ludger Johannes (2004) Clathrin adaptor epsinR is required for retrograde sorting on early endosomal membranes. Dev. Cell 6, 525-538. (abstract) (pdf)
This paper builds on our 2003 epsinR paper by Mills et al, where we found that epsinR overexpression had a trafficking defect in the uptake of TGN46 antibodies. Here we define epsinR as a component of a vesicle budding step in retrograde transport from endosomes to the TGN. This pathway is used for the retrieval of cargo (including TGN38/46 and mannose-6-phosphate receptors) from endosomes and Shiga toxin can also traffic to the TGN via this pathway.

Carlton, J., Bujny, M., Peter, B.J., Oorschot, V.M., Rutherford, A., Mellor, H., Klumperman, J., McMahon, H.T. and Cullen, P.J. (2004) Sorting nexin-1 mediates tubular endosome-to-TGN transport through coincidence sensing of high- curvature membranes and 3-phosphoinositides. Current Biol. Oct 26;14(20):1791-800. (abstract) (pdf) (see also Research Roundup ‘Snx1 hugs the curves’ in JCB 2004 167, p581, pdf) (supplementary material and movies)
Arising from our observations that many sorting nexin proteins have BAR domains (see Peter et al 2004 below) we tested sorting nexin1 for membrane binding and curvature sensing and made appropriate mutations. We then collaborated with Peter Cullen (Bristol University) to test the in vivo significance for protein trafficking in cells. This paper reinforces our previous observation of the necessity for a lipid selective domain and a curvature sensing domain to work in tandem to localize proteins to high curvature on membranes (in this case endosomes) in the cell. This paper also demonstrates that overexpression of this sorting nexin leads to extreme tubulation of endosomes which disrupts all trafficking through this compartments, but an RNAi experiment shows that knock-down of the protein specifically disrupts the trafficking of mannose-6-phosphate receptors to the TGN and thus we propose that this receptor goes though via a tubule transporter which is formed/stabilised by sorting nexin 1.
See also BAR domain web pages

McMahon, H.T. and Mills, I.G. (2004) COP and clathrin-coated vesicle budding: different pathways, common approaches. Current Opinions in Cell Biol. 16, 379-391. (abstract) (pdf)
The remarkable similarities of appendage domains of COP and clathrin adaptors led us to look closely at the overall evolutionary relationships between COP and clathrin-coated vesicles. This review covers cargo recruitment, membrane curvature mechanisms and coat polymerization showing that common paradigms underpin these diverse pathways.
Web pages have been written to accompany this review including movies you can download

Tsuboi, T., McMahon, H.T. and Rutter, G.A. (2004) Mechanisms of dense core vesicle recapture following "kiss and run" ("cavicapture") exocytosis in insulin-secreting cells. J. Biol. Chem. 279 47115-47124. (abstract) (pdf) (Movies)
This paper is the result of a collaboration with Guy Rutter in Bristol University showing the dynamin-dependence of large-dense core vesicle endocytosis. This shows that the not only clathrin-mediated retrieval of vesicles but also the ?kiss and run? form of endocytosis is dynamin dependent. The stabilisation and closure of the fusion pore, through which transmitter is released, required the GTPase activity of dynamin.
See dynamin web pages

Praefcke, GJ.K. and McMahon, H.T. (2004) The Dynamin Superfamily: Universal membrane tubulation and fission molecules? Nature Reviews in Molecular Cell Biology 5, 133-147. (abstract) (pdf)

Here for the first time we define and classify the dynamin ‘Superfamily’ of proteins. All members have GTPase, oligomerisation and membrane targeting domains. The superfamily is subdivided into classical dynamins, dynamin-like proteins, Optic atrophy proteins (OPA1), Mx proteins and GBPs/atlastins. In this review we propose a common mechanism leading to membrane tubulation and/or fission that encompasses all superfamily members. The COS cell on the left is overexpressing a GTPase mutant of dynamin1 (red, T65A-dynaminI) which forms a network of tubules at the end of which we can sometimes see a concentration of transferrin receptors in a fused compartment still accessible to the surface (green,transferrin). The Dynamin web pages were written to accompany this review

see also Significant papers from the past