Sterol-dependent membrane association of the marine sponge-derived bicyclic peptide Theonellamide A as examined by 1H NMR |
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| Affiliation: | 1. Department of Chemistry, Graduate School of Science, Osaka University, Osaka 563-0043, Japan;2. Lipid Active Structure Project, ERATO, Japan Science and Technology Agency, Osaka 560-0043, Japan;3. Department of Chemistry, Graduate School of Sciences, Kyushu University, Fukuoka 819-0395, Japan;4. Division of Bioinformatics and Chemical Genomics, Graduate School of Pharmaceutical Sciences, Kyoto University, Kyoto 606-8501, Japan;5. Advanced Science Institute, RIKEN Center for Sustainable Resource Science, Saitama 351-0198, Japan;6. Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo 113-8657, Japan;1. 1st and 2nd Departments of Radiology, University of Athens, Athens;2. Department of Pathology, University of Athens, Athens;3. Materials Science and Engineering, University of Ioannina, Ioannina;4. Pharmacology & Pharmacotechnology, Biomedical Research Foundation of the Academy of Athens, 4 Soranou Ephessiou Street, Athens 11527, Greece;5. Experimental Surgery, Biomedical Research Foundation of the Academy of Athens, 4 Soranou Ephessiou Street, Athens 11527, Greece;1. Department of Chemistry, Graduate School of Science and Engineering, Tokyo Metropolitan University, 1-1 Minami-Osawa, Hachioji-shi, Tokyo 192-0397, Japan;2. CREST/Japan Science and Technology Agency (JST), 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan;3. Institute of Molecular, Cell and Systems Biology & School of Life Sciences, College of Medical, Veterinary & Life Sciences, Joseph Black Building, University of Glasgow, Glasgow G12 8QQ, UK;4. Institute of Biophysical Chemistry, Center for Biomolecular Magnetic Resonance, Goethe University Frankfurt am Main, Max-von-Laue-Str. 9, 60438 Frankfurt am Main, Germany;5. Frankfurt Institute for Advanced Studies, Ruth-Moufang-Str. 1, 60438 Frankfurt am Main, Germany;1. Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan;2. Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan;3. JST, PRESTO, 3-1 Tanabe-dori, Mizuho-ku, Nagoya 467-8603, Japan;4. Department of Theoretical and Computational Molecular Science, Institute for Molecular Science, National Institutes of Natural Sciences, 38 Nishigo-Naka, Myodaiji, Okazaki, Aichi 444-8585, Japan;5. Department of Structural Molecular Science, The Graduate University for Advanced Studies, Okazaki, Aichi 444-8585, Japan;6. Division of Biomolecular Science, Institute for Molecular Science, National Institutes of Natural Sciences, 5-1 Higashiyama, Myodaiji, Okazaki, Aichi 444-8787, Japan;7. Department of Functional Molecular Science, The Graduate University for Advanced Studies, Okazaki, Aichi 444-8585, Japan;1. Department of Applied Chemistry, Graduate School of Engineering, Osaka University, 2-1 Yamadaoka, Suita, Osaka 565-0871, Japan;2. Interdisciplinary Program for Biomedical Sciences, Institute for Academic Initiatives, Osaka University, 1-1 Yamadaoka, Suita, Osaka 565-0871, Japan;1. Institute of Physics, Kazan Federal University, Kremlevskaya Str., 18, 420008 Kazan, Russia;2. Chemistry Institute, Kazan Federal University, Kremlevskaya Str., 18, 420008 Kazan, Russia;3. Chemistry of Interfaces, Luleå University of Technology, SE-91187 Luleå, Sweden |
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| Abstract: | Theonellamide A (TNM-A) is an antifungal bicyclic dodecapeptide isolated from a marine sponge Theonella sp. Previous studies have shown that TNM-A preferentially binds to 3β-hydroxysterol-containing membranes and disrupts membrane integrity. In this study, several 1H NMR-based experiments were performed to investigate the interaction mode of TNM-A with model membranes. First, the aggregation propensities of TNM-A were examined using diffusion ordered spectroscopy; the results indicate that TNM-A tends to form oligomeric aggregates of 2–9 molecules (depending on peptide concentration) in an aqueous environment, and this aggregation potentially influences the membrane-disrupting activity of the peptide. Subsequently, we measured the 1H NMR spectra of TNM-A with sodium dodecyl sulfate-d25 (SDS-d25) micelles and small dimyristoylphosphatidylcholine (DMPC)-d54/dihexanoylphosphatidylcholine (DHPC)-d22 bicelles in the presence of a paramagnetic quencher Mn2+. These spectra indicate that TNM-A poorly binds to these membrane mimics without sterol and mostly remains in the aqueous media. In contrast, broader 1H signals of TNM-A were observed in 10 mol % cholesterol-containing bicelles, indicating that the peptide efficiently binds to sterol-containing bilayers. The addition of Mn2+ to these bicelles also led to a decrease in the relative intensity and further line-broadening of TNM-A signals, indicating that the peptide stays near the surface of the bilayers. A comparison of the relative signal intensities with those of phospholipids showed that TNM-A resides in the lipid–water interface (close to the C2′ portion of the phospholipid acyl chain). This shallow penetration of TNM-A to lipid bilayers induces an uneven membrane curvature and eventually disrupts membrane integrity. These results shed light on the atomistic mechanism accounting for the membrane-disrupting activity of TNM-A and the important role of cholesterol in its mechanism of action. |
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| Keywords: | Marine sponge Cyclic peptide Cholesterol Membrane curvature |
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