{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,28]],"date-time":"2026-09-28T05:28:14Z","timestamp":1790573294457,"version":"4.1.0"},"reference-count":42,"publisher":"American Chemical Society (ACS)","issue":"35","license":[{"start":{"date-parts":[[2017,7,25]],"date-time":"2017-07-25T00:00:00Z","timestamp":1500940800000},"content-version":"vor","delay-in-days":365,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/100000052","name":"NIH Office of the Director","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100000052","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000062","name":"National Institute of Diabetes and Digestive and Kidney Diseases","doi-asserted-by":"publisher","award":["ZIA DK029047-09"],"award-info":[{"award-number":["ZIA DK029047-09"]}],"id":[{"id":"10.13039\/100000062","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2016,9,6]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>The interaction between \u03b1-synuclein (\u03b1S) protein and lipid membranes is key to its role in synaptic vesicle homeostasis and plays a role in initiating fibril formation, which is implicated in Parkinson\u2019s disease. The natural state of \u03b1S inside the cell is generally believed to be intrinsically disordered, but chemical cross-linking experiments provided evidence of a tetrameric arrangement, which was reported to be rich in \u03b1-helical secondary structure based on circular dichroism (CD). Cross-linking relies on chemical modification of the protein\u2019s Lys C\u03b5 amino groups, commonly by glutaraldehyde, or by disuccinimidyl glutarate (DSG), with the latter agent preferred for cellular assays. We used ultra-high-resolution homonuclear decoupled nuclear magnetic resonance experiments to probe the reactivity of the 15 \u03b1S Lys residues toward N-succinimidyl acetate, effectively half the DSG cross-linker, which results in acetylation of Lys. The intensities of both side chain and backbone amide signals of acetylated Lys residues provide direct information about the reactivity, showing a difference of a factor of 2.5 between the most reactive (K6) and the least reactive (K102) residue. The presence of phospholipid vesicles decreases reactivity of most Lys residues by up to an order of magnitude at high lipid:protein stoichiometries (500:1), but only weakly at low ratios. The decrease in Lys reactivity is found to be impacted by lipid composition, even for vesicles that yield similar \u03b1S CD signatures. Our data provide new insight into the \u03b1S\u2013bilayer interaction, including the pivotal state in which the available lipid surface is limited. Protection of Lys C\u03b5 amino groups by \u03b1S\u2013bilayer interaction will strongly impact quantitative interpretation of DSG cross-linking experiments.<\/jats:p>","DOI":"10.1021\/acs.biochem.6b00637","type":"journal-article","created":{"date-parts":[[2016,7,25]],"date-time":"2016-07-25T13:25:21Z","timestamp":1469453121000},"page":"4949-4959","source":"Crossref","is-referenced-by-count":19,"title":["Nuclear Magnetic Resonance Observation of \u03b1-Synuclein\nMembrane Interaction by Monitoring\nthe Acetylation Reactivity of Its Lysine Side Chains"],"prefix":"10.1021","volume":"55","author":[{"given":"Jung Ho","family":"Lee","sequence":"first","affiliation":[{"name":"National Institutes of Health , , , ,","place":["Bethesda, Maryland, United States, 20892"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jinfa","family":"Ying","sequence":"additional","affiliation":[{"name":"National Institutes of Health , , , ,","place":["Bethesda, Maryland, United States, 20892"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ad","family":"Bax","sequence":"additional","affiliation":[{"name":"National Institutes of Health , , , ,","place":["Bethesda, Maryland, United States, 20892"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2016,7,25]]},"reference":[{"key":"2026090213310622900_cit1","doi-asserted-by":"publisher","first-page":"663","DOI":"10.1002\/mas.20082","article-title":"Chemical cross-linking\nand mass spectrometry to map three-dimensional protein structures\nand protein-protein interactions","volume":"25","author":"Sinz","year":"2006","journal-title":"Mass Spectrom.\nRev."},{"key":"2026090213310622900_cit2","doi-asserted-by":"publisher","first-page":"53","DOI":"10.1016\/0019-2791(69)90178-5","article-title":"Cross-Linking of Proteins\nwith Glutaraldehyde and Its\nUse for Preparation of Immunoadsorbents","volume":"6","author":"Avrameas","year":"1969","journal-title":"Immunochemistry"},{"key":"2026090213310622900_cit3","doi-asserted-by":"publisher","first-page":"6371","DOI":"10.1074\/jbc.M112.403311","article-title":"In Vivo Cross-linking\nReveals Principally Oligomeric Forms of alpha-Synuclein and beta-Synuclein\nin Neurons and Non-neural Cells","volume":"288","author":"Dettmer","year":"2013","journal-title":"J. Biol. Chem."},{"key":"2026090213310622900_cit4","doi-asserted-by":"publisher","first-page":"15345","DOI":"10.1074\/jbc.M111.318949","article-title":"alpha-Synuclein\nin Central Nervous System and from Erythrocytes, Mammalian Cells,\nand Escherichia coli Exists Predominantly as Disordered Monomer","volume":"287","author":"Fauvet","year":"2012","journal-title":"J. Biol. Chem."},{"key":"2026090213310622900_cit5","doi-asserted-by":"publisher","first-page":"107","DOI":"10.1038\/nature10324","article-title":"alpha-Synuclein occurs physiologically\nas a helically folded tetramer that resists aggregation","volume":"477","author":"Bartels","year":"2011","journal-title":"Nature"},{"key":"2026090213310622900_cit6","doi-asserted-by":"publisher","first-page":"7314","DOI":"10.1038\/ncomms8314","article-title":"Parkinson-causing alpha-synuclein missense mutations\nshift native tetramers to monomers as a mechanism for disease initiation","volume":"6","author":"Dettmer","year":"2015","journal-title":"Nat. Commun."},{"key":"2026090213310622900_cit7","doi-asserted-by":"publisher","first-page":"279","DOI":"10.1021\/bi501188a","article-title":"Purification\nof alpha-Synuclein from Human Brain Reveals an Instability of Endogenous\nMultimers as the Protein Approaches Purity","volume":"54","author":"Luth","year":"2015","journal-title":"Biochemistry"},{"key":"2026090213310622900_cit8","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1038\/nature16531","article-title":"Structural\ndisorder\nof monomeric alpha-synuclein persists in mammalian cells","volume":"530","author":"Theillet","year":"2016","journal-title":"Nature"},{"key":"2026090213310622900_cit9","doi-asserted-by":"publisher","first-page":"10251","DOI":"10.1038\/ncomms10251","article-title":"Intracellular repair of oxidation-damaged alpha-synuclein\nfails to target C-terminal modification sites","volume":"7","author":"Binolfi","year":"2016","journal-title":"Nat. Commun."},{"key":"2026090213310622900_cit10","doi-asserted-by":"publisher","first-page":"E4274","DOI":"10.1073\/pnas.1416598111","article-title":"alpha-Synuclein\nassembles into higher-order\nmultimers upon membrane binding to promote SNARE complex formation","volume":"111","author":"Burre","year":"2014","journal-title":"Proc. Natl. Acad. Sci. U. S. A."},{"key":"2026090213310622900_cit11","doi-asserted-by":"publisher","first-page":"17620","DOI":"10.1074\/jbc.M113.476697","article-title":"alpha-Synuclein Oligomers\nwith Broken Helical Conformation\nForm Lipoprotein Nanoparticles","volume":"288","author":"Varkey","year":"2013","journal-title":"J. Biol. Chem."},{"key":"2026090213310622900_cit12","doi-asserted-by":"publisher","first-page":"8516","DOI":"10.1074\/jbc.M115.707968","article-title":"Preparation and Characterization\nof Stable -Synuclein Lipoprotein Particles","volume":"291","author":"Eichmann","year":"2016","journal-title":"J. Biol. 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Chem."},{"key":"2026090213310622900_cit16","doi-asserted-by":"publisher","first-page":"15313","DOI":"10.1074\/jbc.M213128200","article-title":"A broken\nalpha-helix in folded alpha-synuclein","volume":"278","author":"Chandra","year":"2003","journal-title":"J. Biol.\nChem."},{"key":"2026090213310622900_cit17","doi-asserted-by":"publisher","first-page":"21966","DOI":"10.1074\/jbc.M401076200","article-title":"alpha-synuclein has a high affinity\nfor packing defects in a bilayer membrane - A thermodynamics study","volume":"279","author":"Nuscher","year":"2004","journal-title":"J. Biol. 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Biol. Chem."},{"key":"2026090213310622900_cit26","doi-asserted-by":"publisher","first-page":"6661","DOI":"10.1021\/cr400695p","article-title":"Physicochemical Properties of Cells\nand Their Effects\non Intrinsically Disordered Proteins (IDPs)","volume":"114","author":"Theillet","year":"2014","journal-title":"Chem. Rev."},{"key":"2026090213310622900_cit27","doi-asserted-by":"publisher","first-page":"e15801","DOI":"10.1371\/journal.pone.0015801","article-title":"Targeted\nAmino-Terminal Acetylation of Recombinant Proteins in E. coli","volume":"5","author":"Johnson","year":"2010","journal-title":"PLoS One"},{"key":"2026090213310622900_cit28","doi-asserted-by":"publisher","first-page":"97","DOI":"10.1016\/j.jmr.2013.11.006","article-title":"Homonuclear decoupling for enhancing\nresolution and sensitivity in NOE and RDC measurements of peptides\nand proteins","volume":"241","author":"Ying","year":"2014","journal-title":"J. Magn. 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Spectrosc."},{"key":"2026090213310622900_cit32","doi-asserted-by":"publisher","first-page":"572","DOI":"10.1002\/cphc.201402704","article-title":"High Accuracy\nof Karplus Equations for Relating Three-Bond J Couplings to Protein\nBackbone Torsion Angles","volume":"16","author":"Li","year":"2015","journal-title":"ChemPhysChem"},{"key":"2026090213310622900_cit33","doi-asserted-by":"publisher","first-page":"274","DOI":"10.1016\/0022-2364(88)90178-3","article-title":"Iterative\nSchemes for Bilinear Operators;\nApplication to Spin Decoupling","volume":"77","author":"Shaka","year":"1988","journal-title":"J. Magn. Reson."},{"key":"2026090213310622900_cit34","doi-asserted-by":"publisher","first-page":"2130","DOI":"10.1016\/j.jmb.2014.02.024","article-title":"Sequence and Membrane\nDeterminants of the Random Coil-Helix\nTransition of alpha-Synuclein","volume":"426","author":"Lokappa","year":"2014","journal-title":"J. Mol. Biol."},{"key":"2026090213310622900_cit35","doi-asserted-by":"publisher","first-page":"3827","DOI":"10.1038\/ncomms4827","article-title":"Direct observation\nof the three regions in alpha-synuclein that determine its membrane-bound\nbehaviour","volume":"5","author":"Fusco","year":"2014","journal-title":"Nat. Commun."},{"key":"2026090213310622900_cit36","doi-asserted-by":"publisher","first-page":"3788","DOI":"10.1016\/j.febslet.2008.10.009","article-title":"Membrane binding\nof oligomeric alpha-synuclein depends on bilayer charge and packing","volume":"582","author":"van Rooijen","year":"2008","journal-title":"FEBS Lett."},{"key":"2026090213310622900_cit37","doi-asserted-by":"publisher","first-page":"73","DOI":"10.1016\/j.plipres.2015.10.005","article-title":"Interaction\nof alpha-synuclein with biomembranes in Parkinson\u2019s disease\n-role of cardiolipin","volume":"61","author":"Ghio","year":"2016","journal-title":"Prog. Lipid Res."},{"key":"2026090213310622900_cit38","doi-asserted-by":"publisher","first-page":"4692","DOI":"10.1529\/biophysj.105.079251","article-title":"Quantification of alpha-synuclein\nbinding to lipid vesicles using fluorescence correlation spectroscopy","volume":"90","author":"Rhoades","year":"2006","journal-title":"Biophys. J."},{"key":"2026090213310622900_cit39","doi-asserted-by":"publisher","first-page":"1663","DOI":"10.1126\/science.1195227","article-title":"alpha-Synuclein\nPromotes SNARE-Complex Assembly in\nVivo and in Vitro","volume":"329","author":"Burre","year":"2010","journal-title":"Science"},{"key":"2026090213310622900_cit40","doi-asserted-by":"publisher","first-page":"1394","DOI":"10.1016\/j.jmb.2007.11.051","article-title":"alpha-Synuclein selectively\nbinds to anionic phospholipids embedded in liquid-disordered domains","volume":"375","author":"Stockl","year":"2008","journal-title":"J. Mol. Biol."},{"key":"2026090213310622900_cit41","doi-asserted-by":"publisher","first-page":"285","DOI":"10.1007\/s12013-007-0014-9","article-title":"Mechanistic\naspects of Parkinson\u2019s disease: alpha-synuclein and the biomembrane","volume":"47","author":"Beyer","year":"2007","journal-title":"Cell Biochem. Biophys."},{"key":"2026090213310622900_cit42","doi-asserted-by":"publisher","first-page":"5221","DOI":"10.1523\/JNEUROSCI.4650-14.2015","article-title":"Definition of a\nMolecular Pathway\nMediating alpha-Synuclein Neurotoxicity","volume":"35","author":"Burre","year":"2015","journal-title":"J.\nNeurosci."}],"container-title":["Biochemistry"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acs.biochem.6b00637","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/55\/35\/4949\/66941882\/acs.biochem.6b00637.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/bichaw\/article-pdf\/55\/35\/4949\/66941882\/acs.biochem.6b00637.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T17:31:13Z","timestamp":1788370273000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/bichaw\/article\/55\/35\/4949\/1478224\/Nuclear-Magnetic-Resonance-Observation-of"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2016,7,25]]},"references-count":42,"journal-issue":{"issue":"35","published-online":{"date-parts":[[2016,7,25]]},"published-print":{"date-parts":[[2016,9,6]]}},"URL":"https:\/\/doi.org\/10.1021\/acs.biochem.6b00637","relation":{},"ISSN":["0006-2960","1520-4995"],"issn-type":[{"value":"0006-2960","type":"print"},{"value":"1520-4995","type":"electronic"}],"subject":[],"published":{"date-parts":[[2016,7,25]]}}}