{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,30]],"date-time":"2026-07-30T17:10:35Z","timestamp":1785431435464,"version":"3.56.0"},"reference-count":27,"publisher":"National Academy of Sciences","issue":"27","license":[{"start":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T00:00:00Z","timestamp":1782864000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by-nc-nd\/4.0\/"}],"funder":[{"DOI":"10.13039\/100020595","name":"National Science and Technology Council","doi-asserted-by":"publisher","award":["114-2113-M-007-023 114-2123-M-007-001"],"award-info":[{"award-number":["114-2113-M-007-023 114-2123-M-007-001"]}],"id":[{"id":"10.13039\/100020595","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.pnas.org"],"crossmark-restriction":true},"short-container-title":["Proc. Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,7,7]]},"abstract":"<jats:p>\n                    Proton-coupled ion transport is a fundamental chemical process underlying membrane physiology, yet how local electrostatics are transduced into gated Ca\n                    <jats:sup>2+<\/jats:sup>\n                    permeation remains poorly defined. Here, we combine single-channel planar bilayer electrophysiology, nanodisc-based double electron\u2013electron resonance spectroscopy, atomistic modeling, and a nanodisc nano-delivery strategy that enables direct functional insertion of purified membrane proteins into live mammalian cells. Applying this integrated toolkit to the bacterial transmembrane Bax-inhibitor-1\u2013containing motif prototype\n                    <jats:italic toggle=\"yes\">Bs<\/jats:italic>\n                    YetJ, we resolve a hierarchical electrostatic gating mechanism governed by two salt bridges with distinct physical roles. A periplasmic E49\u2013R205 interaction functions as a proton-sensitive latch that drives transmembrane helix 2 displacement and controls opening probability, while a cytoplasmic E182\u2013R15 pair operates as a local electrostatic determinant of Ca\n                    <jats:sup>2+<\/jats:sup>\n                    self-block that tunes conductance and selectivity without large-scale conformational change. Quantitative separation of these effects reveals how protonation reshapes the energy landscape of ion permeation. Live-cell Ca\n                    <jats:sup>2+<\/jats:sup>\n                    imaging following nano-delivery recapitulates this gating logic in a cellular membrane setting. Together, this work establishes dual salt-bridge electrostatics as a chemical principle for graded Ca\n                    <jats:sup>2+<\/jats:sup>\n                    leak and introduces nano-delivery as a powerful platform for connecting molecular electrostatics to cellular ion transport.\n                  <\/jats:p>","DOI":"10.1073\/pnas.2536595123","type":"journal-article","created":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T17:41:20Z","timestamp":1782927680000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":0,"title":["Dual salt bridges govern proton gating and calcium leak in\n                    <i>Bs<\/i>\n                    YetJ across bilayers and live cells"],"prefix":"10.1073","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-9792-5004","authenticated-orcid":false,"given":"Chu-Chun","family":"Cheng","sequence":"first","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00zdnkx70","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, National Tsing Hua University","place":["Hsinchu, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1494-1664","authenticated-orcid":false,"given":"Chieh-Chin","family":"Li","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00zdnkx70","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, National Tsing Hua University","place":["Hsinchu, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yun-Shan","family":"Wang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00zdnkx70","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, National Tsing Hua University","place":["Hsinchu, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2139-3021","authenticated-orcid":false,"given":"Chun-Wei","family":"Lin","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00zdnkx70","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, National Tsing Hua University","place":["Hsinchu, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2101-8918","authenticated-orcid":false,"given":"Yun-Wei","family":"Chiang","sequence":"additional","affiliation":[{"id":[{"id":"https:\/\/ror.org\/00zdnkx70","id-type":"ROR","asserted-by":"publisher"}],"name":"Department of Chemistry, National Tsing Hua University","place":["Hsinchu, Taiwan"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"341","published-online":{"date-parts":[[2026,7,1]]},"reference":[{"key":"e_1_3_4_1_2","doi-asserted-by":"publisher","DOI":"10.1023\/B:APPT.0000025806.71000.1c"},{"key":"e_1_3_4_2_2","doi-asserted-by":"publisher","DOI":"10.1016\/S1097-2765(00)80034-9"},{"key":"e_1_3_4_3_2","doi-asserted-by":"publisher","DOI":"10.1038\/cdd.2011.59"},{"key":"e_1_3_4_4_2","doi-asserted-by":"publisher","DOI":"10.15252\/embr.202254978"},{"key":"e_1_3_4_5_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41388-024-03222-x"},{"key":"e_1_3_4_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2016.12.023"},{"key":"e_1_3_4_7_2","doi-asserted-by":"publisher","DOI":"10.1038\/onc.2014.6"},{"key":"e_1_3_4_8_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M800075200"},{"key":"e_1_3_4_9_2","doi-asserted-by":"publisher","DOI":"10.1126\/sciadv.abe5469"},{"key":"e_1_3_4_10_2","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-17802-4"},{"key":"e_1_3_4_11_2","doi-asserted-by":"publisher","DOI":"10.1186\/s13046-022-02573-1"},{"key":"e_1_3_4_12_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.1252043"},{"key":"e_1_3_4_13_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.2014094117"},{"key":"e_1_3_4_14_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.str.2019.03.003"},{"key":"e_1_3_4_15_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamem.2023.184153"},{"key":"e_1_3_4_16_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacs.1c07371"},{"key":"e_1_3_4_17_2","doi-asserted-by":"publisher","DOI":"10.1021\/jacsau.4c00702"},{"key":"e_1_3_4_18_2","doi-asserted-by":"publisher","DOI":"10.5194\/mr-1-209-2020"},{"key":"e_1_3_4_19_2","doi-asserted-by":"publisher","DOI":"10.1039\/D3CP02569A"},{"key":"e_1_3_4_20_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pcbi.1010834"},{"key":"e_1_3_4_21_2","doi-asserted-by":"publisher","DOI":"10.1093\/nar\/gky309"},{"key":"e_1_3_4_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.1813183115"},{"key":"e_1_3_4_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.bbamcr.2015.03.002"},{"key":"e_1_3_4_24_2","doi-asserted-by":"publisher","DOI":"10.1074\/jbc.M115.637306"},{"key":"e_1_3_4_25_2","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0108097"},{"key":"e_1_3_4_26_2","doi-asserted-by":"publisher","DOI":"10.1016\/j.jmr.2004.10.012"},{"key":"e_1_3_4_27_2","unstructured":"Y.-W. Chiang DEER-derived model of open state BsYetJ. Zenodo. https:\/\/doi.org\/10.5281\/zenodo.20278430. Deposited 19 May 2026."}],"container-title":["Proceedings of the National Academy of Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pnas.org\/doi\/pdf\/10.1073\/pnas.2536595123","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T17:43:41Z","timestamp":1782927821000},"score":1,"resource":{"primary":{"URL":"https:\/\/pnas.org\/doi\/10.1073\/pnas.2536595123"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,1]]},"references-count":27,"journal-issue":{"issue":"27","published-print":{"date-parts":[[2026,7,7]]}},"alternative-id":["10.1073\/pnas.2536595123"],"URL":"https:\/\/doi.org\/10.1073\/pnas.2536595123","relation":{},"ISSN":["0027-8424","1091-6490"],"issn-type":[{"value":"0027-8424","type":"print"},{"value":"1091-6490","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,1]]},"assertion":[{"value":"2025-12-23","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-06-02","order":2,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2026-07-01","order":3,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e2536595123"}}