Abstract
We report on the assembly of tumor necrosis factor receptor 1 (TNF-R1) prior to ligand activation and its ligand-induced reorganization at the cell membrane. We apply single-molecule localization microscopy to obtain quantitative information on receptor cluster sizes and copy numbers. Our data suggest a dimeric pre-assembly of TNF-R1, as well as receptor reorganization toward higher oligomeric states with stable populations comprising three to six TNF-R1. Our experimental results directly serve as input parameters for computational modeling of the ligand–receptor interaction. Simulations corroborate the experimental finding of higher-order oligomeric states. This work is a first demonstration how quantitative, super-resolution and advanced microscopy can be used for systems biology approaches at the single-molecule and single-cell level.
Similar content being viewed by others
References
Annibale P, Vanni S, Scarselli M, Rothlisberger U, Radenovic A (2011) Quantitative photo activated localization microscopy: unraveling the effects of photoblinking. PLoS One 6:e22678. doi:10.1371/journal.pone.0022678
Baaske P, Wienken CJ, Reineck P, Duhr S, Braun D (2010) Optical thermophoresis for quantifying the buffer dependence of aptamer binding. Angew Chem Int Ed Engl 49:2238–2241. doi:10.1002/anie.200903998
Banner DW, D’Arcy A, Janes W, Gentz R, Schoenfeld H-J, Broger C, Loetscher H, Lesslauer W (1993) Crystal structure of the soluble human 55 kd TNF receptor-human TNFβ complex: implications for TNF receptor activation. Cell 73:431–445. doi:10.1016/0092-8674(93)90132-A
Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS, Davidson MW, Lippincott-Schwartz J, Hess HF (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642–1645. doi:10.1126/science.1127344
Boschert V, Krippner-Heidenreich A, Branschädel M, Tepperink J, Aird A, Scheurich P (2010) Single chain TNF derivatives with individually mutated receptor binding sites reveal differential stoichiometry of ligand receptor complex formation for TNFR1 and TNFR2. Cell Signal 22:1088–1096. doi:10.1016/j.cellsig.2010.02.011
Brockhaus M, Schoenfeld HJ, Schlaeger EJ, Hunziker W, Lesslauer W, Loetscher H (1990) Identification of two types of tumor necrosis factor receptors on human cell lines by monoclonal antibodies. Proc Natl Acad Sci 87:3127–3131
Chan FK (2007) Three is better than one: pre-ligand receptor assembly in the regulation of TNF receptor signaling. Cytokine 37:101–107. doi:10.1016/j.cyto.2007.03.005
Chan FK, Chun HJ, Zheng L, Siegel RM, Bui KL, Lenardo MJ (2000) A domain in TNF receptors that mediates ligand-independent receptor assembly and signaling. Science 288:2351–2354. doi:10.1126/science.288.5475.2351
Day ES, Cote SM, Whitty A (2012) Binding efficiency of protein–protein complexes. Biochemistry 51:9124–9136. doi:10.1021/bi301039t
Dietz MS, Fricke F, Krüger CL, Niemann HH, Heilemann M (2013a) Receptor–ligand interactions: binding affinities studied by single-molecule and super-resolution microscopy on intact cells. ChemPhysChem. doi:10.1002/cphc.201300755
Dietz MS, Haße D, Ferraris DM, Göhler A, Niemann HH, Heilemann M (2013b) Single-molecule photobleaching reveals increased MET receptor dimerization upon ligand binding in intact cells. BMC Biophys 6:6. doi:10.1186/2046-1682-6-6
Duhr S, Braun D (2006) Why molecules move along a temperature gradient. Proc Natl Acad Sci 103:19678–19682. doi:10.1073/pnas.0603873103
Eck MJ, Sprang SR (1989) The structure of tumor necrosis factor-alpha at 2.6 A resolution. J Biol Chem 264:17595–17605
Endesfelder U, Malkusch S, Flottmann B, Mondry J, Liguzinski P, Verveer PJ, Heilemann M (2011) Chemically induced photoswitching of fluorescent probes—a general concept for super-resolution microscopy. Molecules 16:3106–3118. doi:10.3390/molecules16043106
Endesfelder U, Malkusch S, Fricke F, Heilemann M (2014) A simple method to estimate the average localization precision of a single-molecule localization microscopy experiment. Histochem Cell Biol. doi:10.1007/s00418-014-1192-3
Gould TJ, Hess ST, Bewersdorf J (2012) Optical nanoscopy: from acquisition to analysis. Annu Rev Biomed Eng 14:231–254. doi:10.1146/annurev-bioeng-071811-150025
Greiner JF-W, Müller J, Zeuner M-T, Hauser S, Seidel T, Klenke C, Grunwald L-M, Schomann T, Widera D, Sudhoff H, Kaltschmidt B, Kaltschmidt C (2013) 1,8-Cineol inhibits nuclear translocation of NF-κB p65 and NF-κB-dependent transcriptional activity. Biochim Biophys Acta 1833:2866–2878. doi:10.1016/j.bbamcr.2013.07.001
Grell M, Douni E, Wajant H, Löhden M, Clauss M, Maxeiner B, Georgopoulos S, Lesslauer W, Kollias G, Pfizenmaier K, Scheurich P (1995) The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell 83:793–802. doi:10.1016/0092-8674(95)90192-2
Heidbreder M, Zander C, Malkusch S, Widera D, Kaltschmidt B, Kaltschmidt C, Nair D, Choquet D, Sibarita J-B, Heilemann M (2012) TNF-α influences the lateral dynamics of TNF receptor I in living cells. Biochim Biophys Acta 1823:1984–1989. doi:10.1016/j.bbamcr.2012.06.026
Henkler F, Behrle E, Dennehy KM, Wicovsky A, Peters N, Warnke C, Pfizenmaier K, Wajant H (2005) The extracellular domains of FasL and Fas are sufficient for the formation of supramolecular FasL–Fas clusters of high stability. J Cell Biol 168:1087–1098. doi:10.1083/jcb.200501048
Hohmann H, Remy R, Brockhaus M, van Loon APGM (1989) Two different cell types have different major receptors for human tumor necrosis factor (TNFalpha). J Biol Chem 264:14927–14934
Hymowitz SG, Christinger HW, Fuh G, Ultsch M, O’Connell M, Kelley RF, Ashkenazi A, de Vos AM (1999) Triggering cell death. Mol Cell 4:563–571. doi:10.1016/S1097-2765(00)80207-5
Jackson TL, Lai R (2004) A mathematical model of receptor-mediated apoptosis: dying to know why FasL is a trimer. Math Biosci Eng 1:325–338. doi:10.3934/mbe.2004.1.325
Jerabek-Willemsen M, Wienken CJ, Braun D, Baaske P, Duhr S (2011) Molecular interaction studies using microscale thermophoresis. Assay Drug Dev Technol 9:342–353. doi:10.1089/adt.2011.0380
Jones E, Oliphant T, Peterson P et al (2001) SciPy: open source scientific tools for Python. http://www.scipy.org/
Krippner-Heidenreich A, Tübing F, Bryde S, Willi S, Zimmermann G, Scheurich P (2002) Control of receptor-induced signaling complex formation by the kinetics of ligand/receptor interaction. J Biol Chem 277:44155–44163. doi:10.1074/jbc.M207399200
Kull FC, Jacobs S, Cuatrecasas P (1985) Cellular receptor for 125I-labeled tumor necrosis factor: specific binding, affinity labeling, and relationship to sensitivity. Proc Natl Acad Sci 82:5756–5760
Lando D, Endesfelder U, Berger H, Subramanian L, Dunne PD, McColl J, Klenerman D, Carr AM, Sauer M, Allshire RC, Heilemann M, Laue ED (2012) Quantitative single-molecule microscopy reveals that CENP-A(Cnp1) deposition occurs during G2 in fission yeast. Open Biol 2:120078. doi:10.1098/rsob.120078
Lee H-W, Lee S-H, Lee H-W, Ryu Y-W, Kwon M-H, Kim Y-S (2005) Homomeric and heteromeric interactions of the extracellular domains of death receptors and death decoy receptors. Biochem Biophys Res Commun 330:1205–1212. doi:10.1016/j.bbrc.2005.03.101
Lee S-H, Shin JY, Lee A, Bustamante C (2012) Counting single photoactivatable fluorescent molecules by photoactivated localization microscopy (PALM). Proc Natl Acad Sci 109:17436–17441. doi:10.1073/pnas.1215175109
Lewis AK, Valley CC, Sachs JN (2012) TNFR1 signaling is associated with backbone conformational changes of receptor dimers consistent with overactivation in the R92Q TRAPS mutant. Biochemistry 51:6545–6555. doi:10.1021/bi3006626
MacEwan DJ (2002) TNF ligands and receptors—a matter of life and death. Br J Pharmacol 135:855–875. doi:10.1038/sj.bjp.0704549
Malkusch S, Endesfelder U, Mondry J, Gelléri M, Verveer PJ, Heilemann M (2012) Coordinate-based colocalization analysis of single-molecule localization microscopy data. Histochem Cell Biol 137:1–10. doi:10.1007/s00418-011-0880-5
Malkusch S, Muranyi W, Müller B, Kräusslich H-G, Heilemann M (2013) Single-molecule coordinate-based analysis of the morphology of HIV-1 assembly sites with near-molecular spatial resolution. Histochem Cell Biol 139:173–179. doi:10.1007/s00418-012-1014-4
Muranyi W, Malkusch S, Müller B, Heilemann M, Kräusslich H-G (2013) Super-resolution microscopy reveals specific recruitment of HIV-1 envelope proteins to viral assembly sites dependent on the envelope C-terminal tail. PLoS Pathog 9:e1003198. doi:10.1371/journal.ppat.1003198
Naismith JH, Devine TQ, Brandhuber BJ, Sprang SR (1995) Crystallographic evidence for dimerization of unliganded tumor necrosis factor receptor. J Biol Chem 270:13303–13307. doi:10.1074/jbc.270.22.13303
Nelson G, Paraoan L, Spiller DG, Wilde GJC, Browne MA, Djali PK, Unitt JF, Sullivan E, Floettmann E, White MRH (2002) Multi-parameter analysis of the kinetics of NF-kappaB signalling and transcription in single living cells. J Cell Sci 115:1137–1148
Ori A, Banterle N, Iskar M, Andrés-Pons A, Escher C, Khanh Bui H, Sparks L, Solis-Mezarino V, Rinner O, Bork P, Lemke EA, Beck M (2013) Cell type-specific nuclear pores: a case in point for context-dependent stoichiometry of molecular machines. Mol Syst Biol 9:648. doi:10.1038/msb.2013.4
Ozsoy HZ, Sivasubramanian N, Wieder ED, Pedersen S, Mann DL (2008) Oxidative stress promotes ligand-independent and enhanced ligand-dependent tumor necrosis factor receptor signaling. J Biol Chem 283:23419–23428. doi:10.1074/jbc.M802967200
Presley JF, Cole NB, Schroer TA, Hirschberg K, Zaal KJ, Lippincott-Schwartz J (1997) ER-to-Golgi transport visualized in living cells. Nature 389:81–85. doi:10.1038/38001
Puchner EM, Walter JM, Kasper R, Huang B, Lim WA (2013) Counting molecules in single organelles with superresolution microscopy allows tracking of the endosome maturation trajectory. Proc Natl Acad Sci 110:16015–16020. doi:10.1073/pnas.1309676110
Richter F, Liebig T, Guenzi E, Herrmann A, Scheurich P, Pfizenmaier K, Kontermann RE (2013) Antagonistic TNF receptor one-specific antibody (ATROSAB): receptor binding and in vitro bioactivity. PLoS One 8:e72156. doi:10.1371/journal.pone.0072156
Schindelin J, Arganda-Carreras I, Frise E, Kaynig V, Longair M, Pietzsch T, Preibisch S, Rueden C, Saalfeld S, Schmid B, Tinevez J-Y, White DJ, Hartenstein V, Eliceiri K, Tomancak P, Cardona A (2012) Fiji: an open-source platform for biological-image analysis. Nat Methods 9:676–682. doi:10.1038/nmeth.2019
Schneider-Brachert W, Tchikov V, Neumeyer J, Jakob M, Winoto-Morbach S, Held-Feindt J, Heinrich M, Merkel O, Ehrenschwender M, Adam D, Mentlein R, Kabelitz D, Schütze S (2004) Compartmentalization of TNF receptor 1 signaling: internalized TNF receptosomes as death signaling vesicles. Immunity 21:415–428. doi:10.1016/j.immuni.2004.08.017
Sengupta P, Jovanovic-Talisman T, Skoko D, Renz M, Veatch SL, Lippincott-Schwartz J (2011) Probing protein heterogeneity in the plasma membrane using PALM and pair correlation analysis. Nat Methods 8:969–975. doi:10.1038/nmeth.1704
Siegel RM, Muppidi JR, Sarker M, Lobito A, Jen M, Martin D, Straus SE, Lenardo MJ (2004) SPOTS: signaling protein oligomeric transduction structures are early mediators of death receptor-induced apoptosis at the plasma membrane. J Cell Biol 167:735–744. doi:10.1083/jcb.200406101
Sneddon MW, Faeder JR, Emonet T (2011) Efficient modeling, simulation and coarse-graining of biological complexity with NFsim. Nat Methods 8:177–183. doi:10.1038/nmeth.1546
Valley CC, Lewis AK, Mudaliar DJ, Perlmutter JD, Braun AR, Karim CB, Thomas DD, Brody JR, Sachs JN (2012) Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) induces death receptor 5 networks that are highly organized. J Biol Chem 287:21265–21278. doi:10.1074/jbc.M111.306480
Vandenabeele P, Galluzzi L, Vanden Berghe T, Kroemer G (2010) Molecular mechanisms of necroptosis: an ordered cellular explosion. Nat Rev Mol Cell Biol 11:700–714. doi:10.1038/nrm2970
Wajant H, Scheurich P (2011) TNFR1-induced activation of the classical NF-κB pathway. FEBS J 278:862–876. doi:10.1111/j.1742-4658.2011.08015.x
Widera D, Mikenberg I, Elvers M, Kaltschmidt C, Kaltschmidt B (2006) Tumor necrosis factor alpha triggers proliferation of adult neural stem cells via IKK/NF-kappaB signaling. BMC Neurosci 7:64. doi:10.1186/1471-2202-7-64
Winkel C, Neumann S, Surulescu C, Scheurich P (2012) A minimal mathematical model for the initial molecular interactions of death receptor signalling. Math Biosci Eng 9:663–683. doi:10.3934/mbe.2012.9.663
Wolter S, Löschberger A, Holm T, Aufmkolk S, Dabauvalle M, van de Linde S, Sauer M (2012) rapidSTORM: accurate, fast open-source software for localization microscopy. Nat Methods 9:1040–1041. doi:10.1038/nmeth.2224
Yoshida A, Kohchi C, Inagawa H, Nishizawa T, Hori H, Soma G (2006) A soluble 17 kDa tumour necrosis factor (TNF) mutein, TNF-SAM2, with membrane-bound TNF-like biological characteristics. Anticancer Res 26:4003–4008
Zagouras P, Rose J (1993) Dynamic equilibrium between vesicular stomatitis virus glycoprotein monomers and trimers in the Golgi and at the cell surface. J Virol 67:7533–7538
Acknowledgments
We thank Markus Braner for help with microscale thermophoresis measurements. M. Heilemann, F. Fricke and S. Malkusch acknowledge the financial support by the German Ministry of Education and Research (BMBF, Grant 336314) and the cluster of excellence “Macromolecular Complexes” (CEF, DFG cluster of excellence (EXC 115)). T. Dandekar and G. Wangorsch acknowledge the financial support by the German Science Foundation (DFG, Da 208/12-1 and SFB688/A2). D. Widera, C. Kaltschmidt, B. Kaltschmidt and J. Greiner were supported by the BMBF and Cassella Med, Cologne.
Author information
Authors and Affiliations
Corresponding author
Additional information
Franziska Fricke and Sebastian Malkusch have contributed equally.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Fricke, F., Malkusch, S., Wangorsch, G. et al. Quantitative single-molecule localization microscopy combined with rule-based modeling reveals ligand-induced TNF-R1 reorganization toward higher-order oligomers. Histochem Cell Biol 142, 91–101 (2014). https://doi.org/10.1007/s00418-014-1195-0
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00418-014-1195-0