{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,9]],"date-time":"2026-01-09T02:01:33Z","timestamp":1767924093938,"version":"3.49.0"},"reference-count":52,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,6,21]],"date-time":"2017-06-21T00:00:00Z","timestamp":1498003200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,6,21]],"date-time":"2017-06-21T00:00:00Z","timestamp":1498003200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Synthetic systems are widely used to unveil the molecular mechanisms of complex cellular events. Artificial membranes are key examples of models employed to address lipid-lipid and lipid-protein interactions. In this work, we developed a new synthetic system that more closely resembles the lysosome \u2013 the lysosome-mimicking vesicles (LMVs) \u2013 displaying stable acid-to-neutral pH gradient across the membrane. To evaluate the advantages of this synthetic system, we assessed the distinct effects of sphingosine (Sph) accumulation in membrane structure and biophysical properties of standard liposomes (no pH gradient) and in LMVs with lipid composition tuned to mimic physiological- or NPC1-like lysosomes. Ternary 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC)\/Sphingomyelin (SM)\/Cholesterol (Chol) mixtures with, respectively, low and high Chol\/SM levels were prepared. The effect of Sph on membrane permeability and biophysical properties was evaluated by fluorescence spectroscopy, electrophoretic and dynamic light scattering. The results showed that overall Sph has the ability to cause a shift in vesicle surface charge, increase membrane order and promote a rapid increase in membrane permeability. These effects are enhanced in NPC1- LMVs. The results suggest that lysosomal accumulation of these lipids, as observed under pathological conditions, might significantly affect lysosomal membrane structure and integrity, and therefore contribute to the impairment of cell function.<\/jats:p>","DOI":"10.1038\/s41598-017-04125-6","type":"journal-article","created":{"date-parts":[[2017,6,15]],"date-time":"2017-06-15T10:04:40Z","timestamp":1497521080000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":24,"title":["Development of lysosome-mimicking vesicles to study the effect of abnormal accumulation of sphingosine on membrane properties"],"prefix":"10.1038","volume":"7","author":[{"given":"Ana C.","family":"Carreira","sequence":"first","affiliation":[]},{"given":"Rodrigo F. M.","family":"de Almeida","sequence":"additional","affiliation":[]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1311-0770","authenticated-orcid":false,"given":"Liana C.","family":"Silva","sequence":"additional","affiliation":[]}],"member":"297","published-online":{"date-parts":[[2017,6,21]]},"reference":[{"key":"4125_CR1","doi-asserted-by":"publisher","first-page":"1621","DOI":"10.1194\/jlr.R800012-JLR200","volume":"49","author":"ST Pruett","year":"2008","unstructured":"Pruett, S. T. et al. Biodiversity of sphingoid bases (\u201csphingosines\u201d) and related amino alcohols. J Lipid Res 49, 1621\u20131639 (2008).","journal-title":"J Lipid Res"},{"key":"4125_CR2","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1007\/978-1-4419-6741-1_1","volume":"688","author":"CR Gault","year":"2010","unstructured":"Gault, C. R., Obeid, L. M. & Hannun, Y. A. An overview of sphingolipid metabolism: from synthesis to breakdown. Adv Exp Med Biol 688, 1\u201323 (2010).","journal-title":"Adv Exp Med Biol"},{"key":"4125_CR3","doi-asserted-by":"publisher","first-page":"419","DOI":"10.1111\/j.1600-0854.2010.01032.x","volume":"11","author":"E Lloyd-Evans","year":"2010","unstructured":"Lloyd-Evans, E. & Platt, F. M. Lipids on trial: the search for the offending metabolite in Niemann-Pick type C disease. Traffic 11, 419\u2013428 (2010).","journal-title":"Traffic"},{"key":"4125_CR4","doi-asserted-by":"publisher","first-page":"1010","DOI":"10.1016\/j.cellsig.2007.12.006","volume":"20","author":"K Kitatani","year":"2008","unstructured":"Kitatani, K., Idkowiak-Baldys, J. & Hannun, Y. A. The sphingolipid salvage pathway in ceramide metabolism and signaling. Cell signal 20, 1010\u20131018 (2008).","journal-title":"Cell signal"},{"key":"4125_CR5","doi-asserted-by":"publisher","first-page":"1234","DOI":"10.1111\/j.1600-0854.2012.01379.x","volume":"13","author":"T Blom","year":"2012","unstructured":"Blom, T., Li, Z., Bittman, R., Somerharju, P. & Ikonen, E. Tracking sphingosine metabolism and transport in sphingolipidoses: NPC1 deficiency as a test case. Traffic 13, 1234\u20131243 (2012).","journal-title":"Traffic"},{"key":"4125_CR6","doi-asserted-by":"crossref","first-page":"76","DOI":"10.1016\/S0021-9258(19)40197-X","volume":"265","author":"H Zhang","year":"1990","unstructured":"Zhang, H., Buckley, N. E., Gibson, K. & Spiegel, S. Sphingosine stimulates cellular proliferation via a protein kinase C-independent pathway. J Biol Chem 265, 76\u201381 (1990).","journal-title":"J Biol Chem"},{"key":"4125_CR7","doi-asserted-by":"publisher","first-page":"22","DOI":"10.1016\/j.pmu.2014.03.003","volume":"3","author":"T Kanno","year":"2014","unstructured":"Kanno, T., Gotoh, A. & Nishizaki, T. Sphingosine arrests the cell cycle and induces apoptosis by targeting sphingosine-dependent protein kinase and protein kinase C\u03b4 in vitro. Personalized Medicine Universe 3, 22\u201327 (2014).","journal-title":"Personalized Medicine Universe"},{"key":"4125_CR8","doi-asserted-by":"publisher","first-page":"358","DOI":"10.1002\/(SICI)1097-0215(19960503)66:3<358::AID-IJC16>3.0.CO;2-7","volume":"66","author":"EA Sweeney","year":"1996","unstructured":"Sweeney, E. A. et al. Sphingosine and its methylated derivative N,N-dimethylsphingosine (DMS) induce apoptosis in a variety of human cancer cell lines. Int J Cancer 66, 358\u2013366 (1996).","journal-title":"Int J Cancer"},{"key":"4125_CR9","doi-asserted-by":"crossref","first-page":"12604","DOI":"10.1016\/S0021-9258(18)67133-9","volume":"261","author":"YA Hannun","year":"1986","unstructured":"Hannun, Y. A., Loomis, C. R., Merrill, A. H. Jr. & Bell, R. M. Sphingosine inhibition of protein kinase C activity and of phorbol dibutyrate binding in vitro and in human platelets. J Biol Chem 261, 12604\u201312609 (1986).","journal-title":"J Biol Chem"},{"key":"4125_CR10","doi-asserted-by":"publisher","first-page":"18108","DOI":"10.1074\/jbc.M909663199","volume":"275","author":"CC King","year":"2000","unstructured":"King, C. C. et al. Sphingosine is a novel activator of 3-phosphoinositide-dependent kinase 1. J Biol Chem 275, 18108\u201318113 (2000).","journal-title":"J Biol Chem"},{"key":"4125_CR11","doi-asserted-by":"publisher","first-page":"26011","DOI":"10.1074\/jbc.M409081200","volume":"280","author":"Y Ma","year":"2005","unstructured":"Ma, Y. et al. Sphingosine activates protein kinase A type II by a novel cAMP-independent mechanism. J Biol Chem 280, 26011\u201326017 (2005).","journal-title":"J Biol Chem"},{"key":"4125_CR12","doi-asserted-by":"crossref","first-page":"21773","DOI":"10.1016\/S0021-9258(18)54703-7","volume":"266","author":"OB McDonald","year":"1991","unstructured":"McDonald, O. B., Hannun, Y. A., Reynolds, C. H. & Sahyoun, N. Activation of casein kinase II by sphingosine. J Biol Chem 266, 21773\u201321776 (1991).","journal-title":"J Biol Chem"},{"key":"4125_CR13","doi-asserted-by":"publisher","first-page":"21834","DOI":"10.1074\/jbc.273.34.21834","volume":"273","author":"T Megidish","year":"1998","unstructured":"Megidish, T., Cooper, J., Zhang, L., Fu, H. & Hakomori, S. A novel sphingosine-dependent protein kinase (SDK1) specifically phosphorylates certain isoforms of 14-3-3 protein. J Biol Chem 273, 21834\u201321845 (1998).","journal-title":"J Biol Chem"},{"key":"4125_CR14","doi-asserted-by":"publisher","first-page":"217","DOI":"10.1080\/096876899294544","volume":"16","author":"JE Johnson","year":"1999","unstructured":"Johnson, J. E. & Cornell, R. B. Amphitropic proteins: regulation by reversible membrane interactions (review). Mol Membr Biol 16, 217\u2013235 (1999).","journal-title":"Mol Membr Biol"},{"key":"4125_CR15","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1007\/BF00768840","volume":"23","author":"A Merrill Jr.","year":"1991","unstructured":"Merrill, A. Jr. Cell regulation by sphingosine and more complex sphingolipids. J Bioenerg Biomembr\u00a023, 83\u2013104 (1991).","journal-title":"Journal of Bioenergetics and Biomembranes"},{"key":"4125_CR16","doi-asserted-by":"publisher","first-page":"1247","DOI":"10.1038\/nm.1876","volume":"14","author":"E Lloyd-Evans","year":"2008","unstructured":"Lloyd-Evans, E. et al. Niemann-Pick disease type C1 is a sphingosine storage disease that causes deregulation of lysosomal calcium. Nat Med 14, 1247\u20131255 (2008).","journal-title":"Nat Med"},{"key":"4125_CR17","doi-asserted-by":"publisher","first-page":"723","DOI":"10.1083\/jcb.201208152","volume":"199","author":"FM Platt","year":"2012","unstructured":"Platt, F. M., Boland, B. & van der Spoel, A. C. Lysosomal storage disorders: The cellular impact of lysosomal dysfunction. J Cell Biol 199, 723\u2013734 (2012).","journal-title":"J Cell Biol"},{"key":"4125_CR18","doi-asserted-by":"publisher","first-page":"2577","DOI":"10.1016\/j.bpj.2014.04.038","volume":"106","author":"N Jimenez-Rojo","year":"2014","unstructured":"Jimenez-Rojo, N. et al. Membrane permeabilization induced by sphingosine: effect of negatively charged lipids. Biophys J 106, 2577\u20132584 (2014).","journal-title":"Biophys J"},{"key":"4125_CR19","doi-asserted-by":"publisher","first-page":"1460","DOI":"10.1042\/BST20140145","volume":"42","author":"AM Villamil Giraldo","year":"2014","unstructured":"Villamil Giraldo, A. M., Appelqvist, H., Ederth, T. & Ollinger, K. Lysosomotropic agents: impact on lysosomal membrane permeabilization and cell death. Biochem Soc Trans 42, 1460\u20131464 (2014).","journal-title":"Biochem Soc Trans"},{"key":"4125_CR20","doi-asserted-by":"publisher","first-page":"4085","DOI":"10.1529\/biophysj.105.076471","volume":"90","author":"FX Contreras","year":"2006","unstructured":"Contreras, F. X., Sot, J., Alonso, A. & Go\u00f1i, F. M. Sphingosine Increases the Permeability of Model and Cell Membranes. Biophys J 90, 4085\u20134092 (2006).","journal-title":"Biophys J"},{"key":"4125_CR21","doi-asserted-by":"publisher","first-page":"502","DOI":"10.1016\/j.jcis.2010.07.022","volume":"350","author":"R Georgieva","year":"2010","unstructured":"Georgieva, R., Koumanov, K., Momchilova, A., Tessier, C. & Staneva, G. Effect of sphingosine on domain morphology in giant vesicles. J Colloid Interface Sci 350, 502\u2013510 (2010).","journal-title":"J Colloid Interface Sci"},{"key":"4125_CR22","doi-asserted-by":"publisher","first-page":"123","DOI":"10.1016\/0009-3084(93)90037-4","volume":"66","author":"A Koiv","year":"1993","unstructured":"Koiv, A., Mustonen, P. & Kinnunen, P. K. Influence of sphingosine on the thermal phase behaviour of neutral and acidic phospholipid liposomes. Chem Phys Lipids 66, 123\u2013134 (1993).","journal-title":"Chem Phys Lipids"},{"key":"4125_CR23","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/0005-2736(93)90269-6","volume":"1153","author":"F Lopez-Garcia","year":"1993","unstructured":"Lopez-Garcia, F., Micol, V., Villalain, J. & Gomez-Fernandez, J. C. Interaction of sphingosine and stearylamine with phosphatidylserine as studied by DSC and NMR. Biochim Biophys Acta 1153, 1\u20138 (1993).","journal-title":"Biochim Biophys Acta"},{"key":"4125_CR24","doi-asserted-by":"publisher","first-page":"281","DOI":"10.1016\/0005-2736(94)90310-7","volume":"1194","author":"F Lopez-Garcia","year":"1994","unstructured":"Lopez-Garcia, F., Villalain, J. & Gomez-Fernandez, J. C. A phase behavior study of mixtures of sphingosine with zwitterionic phospholipids. Biochim Biophys Acta 1194, 281\u2013288 (1994).","journal-title":"Biochim Biophys Acta"},{"key":"4125_CR25","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1016\/0005-2736(95)00059-C","volume":"1236","author":"F Lopez-Garcia","year":"1995","unstructured":"Lopez-Garcia, F., Villalain, J. & Gomez-Fernandez, J. C. Effect of sphingosine and stearylamine on the interaction of phosphatidylserine with calcium. A study using DSC, FT-IR and 45Ca(2+)-binding. Biochim Biophys Acta 1236, 279\u2013288 (1995).","journal-title":"Biochim Biophys Acta"},{"key":"4125_CR26","doi-asserted-by":"publisher","first-page":"4858","DOI":"10.1021\/jp501167f","volume":"118","author":"E Zupancic","year":"2014","unstructured":"Zupancic, E., Carreira, A. C., de Almeida, R. F. & Silva, L. C. Biophysical implications of sphingosine accumulation in membrane properties at neutral and acidic pH. J Phys Chem B 118, 4858\u20134866 (2014).","journal-title":"J Phys Chem B"},{"key":"4125_CR27","doi-asserted-by":"publisher","first-page":"13956","DOI":"10.1021\/la5039816","volume":"30","author":"C Watanabe","year":"2014","unstructured":"Watanabe, C., Puff, N., Staneva, G., Seigneuret, M. & Angelova, M. I. Antagonism and Synergy of Single Chain Sphingolipids Sphingosine and Sphingosine-1-phosphate toward Lipid Bilayer Properties. Consequences for Their Role as Cell Fate Regulators. Langmuir 30, 13956\u201313963 (2014).","journal-title":"Langmuir"},{"key":"4125_CR28","doi-asserted-by":"publisher","first-page":"3138","DOI":"10.1021\/bi00434a004","volume":"28","author":"AH Merrill Jr.","year":"1989","unstructured":"Merrill, A. H. Jr. et al. Structural requirements for long-chain (sphingoid) base inhibition of protein kinase C in vitro and for the cellular effects of these compounds. Biochemistry 28, 3138\u20133145 (1989).","journal-title":"Biochemistry"},{"key":"4125_CR29","doi-asserted-by":"crossref","unstructured":"Sasaki, H., Arai, H., Cocco, M. J. & White, S. H. pH dependence of sphingosine aggregation. Biophys J 96, 2727\u20132733 (2009).","DOI":"10.1016\/j.bpj.2008.12.3926"},{"key":"4125_CR30","doi-asserted-by":"crossref","first-page":"10616","DOI":"10.7554\/eLife.10616","volume":"27","author":"D Hoglinger","year":"2015","unstructured":"Hoglinger, D. et al. Intracellular sphingosine releases calcium from lysosomes. Elife 27, 10616 (2015).","journal-title":"Elife"},{"key":"4125_CR31","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1007\/s10863-005-6632-2","volume":"37","author":"LJ Siskind","year":"2005","unstructured":"Siskind, L. J., Fluss, S., Bui, M. & Colombini, M. Sphingosine forms channels in membranes that differ greatly from those formed by ceramide. J Bioenerg Biomembr 37, 227\u2013236 (2005).","journal-title":"J Bioenerg Biomembr"},{"key":"4125_CR32","first-page":"674","volume":"1793","author":"H Schulze","year":"2009","unstructured":"Schulze, H., Kolter, T. & Sandhoff, K. Principles of lysosomal membrane degradation: Cellular topology and biochemistry of lysosomal lipid degradation. BBA-Mol Cell Res 1793, 674\u2013683 (2009).","journal-title":"(BBA)-Mol Cell Res"},{"key":"4125_CR33","doi-asserted-by":"publisher","first-page":"222","DOI":"10.1034\/j.1600-0854.2003.00072.x","volume":"4","author":"W Mobius","year":"2003","unstructured":"Mobius, W. et al. Recycling compartments and the internal vesicles of multivesicular bodies harbor most of the cholesterol found in the endocytic pathway. Traffic 4, 222\u2013231 (2003).","journal-title":"Traffic"},{"key":"4125_CR34","doi-asserted-by":"publisher","first-page":"2406","DOI":"10.1016\/S0006-3495(03)74664-5","volume":"85","author":"RFM de Almeida","year":"2003","unstructured":"de Almeida, R. F. M., Fedorov, A. & Prieto, M. Sphingomyelin\/Phosphatidylcholine\/Cholesterol Phase Diagram: Boundaries and Composition of Lipid Rafts. Biophys J 85, 2406\u20132416 (2003).","journal-title":"Biophys J"},{"key":"4125_CR35","doi-asserted-by":"publisher","first-page":"3264","DOI":"10.1529\/biophysj.104.044883","volume":"87","author":"A Coutinho","year":"2004","unstructured":"Coutinho, A., Silva, L., Fedorov, A. & Prieto, M. Cholesterol and ergosterol influence nystatin surface aggregation: relation to pore formation. Biophys J 87, 3264\u20133276 (2004).","journal-title":"Biophys J"},{"key":"4125_CR36","doi-asserted-by":"publisher","first-page":"1639","DOI":"10.1529\/biophysj.107.107714","volume":"93","author":"BM Castro","year":"2007","unstructured":"Castro, B. M., de Almeida, R. F., Silva, L. C., Fedorov, A. & Prieto, M. Formation of ceramide\/sphingomyelin gel domains in the presence of an unsaturated phospholipid: a quantitative multiprobe approach. Biophys J 93, 1639\u20131650 (2007).","journal-title":"Biophys J"},{"key":"4125_CR37","doi-asserted-by":"publisher","first-page":"5260","DOI":"10.1073\/pnas.1010750108","volume":"108","author":"DG Isom","year":"2011","unstructured":"Isom, D. G., Casta\u00f1eda, C. A., Cannon, B. R. & Garc\u00eda-Moreno, E. B. Large shifts in pKa values of lysine residues buried inside a protein. P Natl Acad Sci 108, 5260\u20135265 (2011).","journal-title":"P Natl Acad Sci"},{"key":"4125_CR38","first-page":"1","volume":"1","author":"DK Hincha","year":"2003","unstructured":"Hincha, D. K. Effects of calcium-induced aggregation on the physical stability of liposomes containing plant glycolipids. Biochim Biophys Acta 1, 1\u20132 (2003).","journal-title":"Biochim Biophys Acta"},{"key":"4125_CR39","doi-asserted-by":"crossref","unstructured":"Schwille, P. In The Minimal Cell: The Biophysics of Cell Compartment and the Origin of Cell Functionality (eds Pier Luigi Luisi & Pasquale Stano) 231\u2013253 (Springer Netherlands, 2011).","DOI":"10.1007\/978-90-481-9944-0_13"},{"key":"4125_CR40","doi-asserted-by":"publisher","first-page":"223","DOI":"10.1080\/10409230903074549","volume":"44","author":"P Schwille","year":"2009","unstructured":"Schwille, P. & Diez, S. Synthetic biology of minimal systems. Crit Rev Biochem Mol Biol 44, 223\u2013242 (2009).","journal-title":"Crit Rev Biochem Mol Biol"},{"key":"4125_CR41","doi-asserted-by":"publisher","first-page":"20150038","DOI":"10.1098\/rsfs.2015.0038","volume":"5","author":"TJ Lagny","year":"2015","unstructured":"Lagny, T. J. & Bassereau, P. Bioinspired membrane-based systems for a physical approach of cell organization and dynamics: usefulness and limitations. Interface Focus 5, 20150038 (2015).","journal-title":"Interface Focus"},{"key":"4125_CR42","doi-asserted-by":"publisher","first-page":"5005","DOI":"10.1038\/srep05005","volume":"4","author":"A Magarkar","year":"2014","unstructured":"Magarkar, A. et al. Cholesterol level affects surface charge of lipid membranes in saline solution. Sci Rep 4, 5005 (2014).","journal-title":"Sci Rep"},{"key":"4125_CR43","doi-asserted-by":"publisher","first-page":"1003","DOI":"10.1248\/cpb.41.1003","volume":"41","author":"N Hazemoto","year":"1993","unstructured":"Hazemoto, N. et al. Effect of phosphatidylcholine and cholesterol on pH-sensitive liposomes. Chem Pharm Bull 41, 1003\u20131006 (1993).","journal-title":"Chem Pharm Bull"},{"key":"4125_CR44","doi-asserted-by":"publisher","first-page":"12036","DOI":"10.1074\/jbc.M302531200","volume":"279","author":"H Deguchi","year":"2004","unstructured":"Deguchi, H., Yegneswaran, S. & Griffin, J. H. Sphingolipids as bioactive regulators of thrombin generation. J Biol Chem 279, 12036\u201312042 (2004).","journal-title":"J Biol Chem"},{"key":"4125_CR45","doi-asserted-by":"crossref","unstructured":"Mateo, C. R., G\u00f3mez, J., Villala\u00edn, J. & Ros, J. M. G. Protein-Lipid Interactions: New Approaches and Emerging Concepts. (Springer Berlin Heidelberg, 2006).","DOI":"10.1007\/3-540-28435-4"},{"key":"4125_CR46","doi-asserted-by":"publisher","first-page":"3477","DOI":"10.1073\/pnas.72.9.3477","volume":"72","author":"PL Yeagle","year":"1975","unstructured":"Yeagle, P. L., Hutton, W. C., Huang, C. H. & Martin, R. B. Headgroup conformation and lipid\u2013cholesterol association in phosphatidylcholine vesicles: a 31P(1H) nuclear Overhauser effect study. P Natl Acad Sci 72, 3477\u20133481 (1975).","journal-title":"P Natl Acad Sci"},{"key":"4125_CR47","doi-asserted-by":"publisher","first-page":"12627","DOI":"10.1021\/la503086a","volume":"30","author":"JT Marqu\u00eas","year":"2014","unstructured":"Marqu\u00eas, J. T., Viana, A. S. & de Almeida, R. F. M. A Biomimetic Platform to Study the Interactions of Bioelectroactive Molecules with Lipid Nanodomains. Langmuir 30, 12627\u201312637 (2014).","journal-title":"Langmuir"},{"key":"4125_CR48","doi-asserted-by":"publisher","first-page":"5043","DOI":"10.1074\/jbc.M110.154435","volume":"286","author":"F Aresta-Branco","year":"2011","unstructured":"Aresta-Branco, F. et al. Gel domains in the plasma membrane of Saccharomyces cerevisiae: highly ordered, ergosterol-free, and sphingolipid-enriched lipid rafts. J Biol Chem 286, 5043\u20135054 (2011).","journal-title":"J Biol Chem"},{"key":"4125_CR49","doi-asserted-by":"publisher","first-page":"3099","DOI":"10.1021\/bi00334a005","volume":"24","author":"H Ellens","year":"1985","unstructured":"Ellens, H., Bentz, J. & Szoka, F. C. Proton- and calcium-induced fusion and destabilization of liposomes. Biochemistry 24, 3099\u20133106 (1985).","journal-title":"Biochemistry"},{"key":"4125_CR50","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1007\/BF02668129","volume":"1","author":"G Rouser","year":"1966","unstructured":"Rouser, G., Siakotos, A. N. & Fleischer, S. Quantitative analysis of phospholipids by thin-layer chromatography and phosphorus analysis of spots. Lipids 1, 85\u201386 (1966).","journal-title":"Lipids"},{"key":"4125_CR51","doi-asserted-by":"crossref","unstructured":"Lakowicz, J. R. Principles of Fluorescence Spectroscopy. (Springer, 2006).","DOI":"10.1007\/978-0-387-46312-4"},{"key":"4125_CR52","doi-asserted-by":"publisher","first-page":"3156","DOI":"10.1073\/pnas.92.8.3156","volume":"92","author":"CC Overly","year":"1995","unstructured":"Overly, C. C., Lee, K. D., Berthiaume, E. & Hollenbeck, P. J. Quantitative measurement of intraorganelle pH in the endosomal-lysosomal pathway in neurons by using ratiometric imaging with pyranine. P Natl Acad Sci USA 92, 3156\u20133160 (1995).","journal-title":"P Natl Acad Sci USA"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04125-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04125-6","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04125-6.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,8,24]],"date-time":"2023-08-24T01:31:26Z","timestamp":1692840686000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-04125-6"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,6,21]]},"references-count":52,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["4125"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-04125-6","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,6,21]]},"assertion":[{"value":"24 June 2016","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"10 May 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"21 June 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"3949"}}