{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T13:23:14Z","timestamp":1787404994723,"version":"build-2736575974"},"reference-count":16,"publisher":"American Chemical Society (ACS)","issue":"8","license":[{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-029"},{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-037"},{"start":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T00:00:00Z","timestamp":1784678400000},"content-version":"stm-asf","delay-in-days":0,"URL":"https:\/\/doi.org\/10.15223\/policy-045"}],"funder":[{"DOI":"10.13039\/100000165","name":"Division of Chemistry","doi-asserted-by":"publisher","award":["CHE-2235835"],"award-info":[{"award-number":["CHE-2235835"]}],"id":[{"id":"10.13039\/100000165","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100007270","name":"University of Michigan","doi-asserted-by":"publisher","award":["NA"],"award-info":[{"award-number":["NA"]}],"id":[{"id":"10.13039\/100007270","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100023581","name":"National Science Foundation Graduate Research Fellowship Program","doi-asserted-by":"publisher","award":["DGE 1841052"],"award-info":[{"award-number":["DGE 1841052"]}],"id":[{"id":"10.13039\/100023581","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2026,8,21]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Protease cleavage is a crucial process, associated with cellular signaling and viral infection. Here, we present a novel sensing motif based on a proximity-dependent biotin ligase, TurboID. The sensor, Biotin Annotating Genetically Encoded Ligase-based Sensor (BAGELS), was created by engineering TurboID such that it is only active following protease activity, linking cleavage events to the biotinylation of proximal proteins. BAGELS produces a signal-to-background ratio of approximately 48 when tested with and without protease. This reporter represents the first protease activity sensor with biotin ligase readout in neurons and HEK293T cells. We demonstrate the reporter\u2019s use for characterization of protease inhibitors and multiplexed imaging with fluorogenic sensors. Overall, this sensor offers a versatile, enzymatically amplified biotin-based readout compatible with biotin-based detection techniques.<\/jats:p>","DOI":"10.1021\/acschembio.6c00334","type":"journal-article","created":{"date-parts":[[2026,7,22]],"date-time":"2026-07-22T11:03:02Z","timestamp":1784718182000},"page":"1952-1957","source":"Crossref","is-referenced-by-count":0,"title":["Biotin Ligase-Based\nReporter for Detecting Protease\nActivity"],"prefix":"10.1021","volume":"21","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-7424-8197","authenticated-orcid":true,"given":"Jennifer","family":"Sescil","sequence":"first","affiliation":[{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]},{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Isabel","family":"Solowiej","sequence":"additional","affiliation":[{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]},{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7290-6800","authenticated-orcid":true,"given":"Guanwei","family":"Zhou","sequence":"additional","affiliation":[{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]},{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Tae","family":"Heo","sequence":"additional","affiliation":[{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Emma","family":"Fu","sequence":"additional","affiliation":[{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6025-9848","authenticated-orcid":true,"given":"Wenjing","family":"Wang","sequence":"additional","affiliation":[{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]},{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]},{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109"]},{"name":"University of Michigan , , , ,","place":["Ann Arbor, Michigan, United States, 48109-1382"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2026,7,22]]},"reference":[{"issue":"45","key":"2026082208551690400_cit1","doi-asserted-by":"publisher","first-page":"30433","DOI":"10.1074\/jbc.R800035200","article-title":"Proteases: multifunctional enzymes in life and disease","volume":"283","author":"Lopez-Otin","year":"2008","journal-title":"J. Biol. Chem."},{"issue":"5","key":"2026082208551690400_cit2","doi-asserted-by":"publisher","first-page":"1643","DOI":"10.1074\/jbc.TM118.004156","article-title":"Proteases:\nHistory, discovery, and roles in health and disease","volume":"294","author":"Bond","year":"2019","journal-title":"J. Biol. Chem."},{"key":"2026082208551690400_cit3","doi-asserted-by":"publisher","first-page":"101144","DOI":"10.1016\/j.mam.2022.101144","article-title":"Diagnostic\nand therapeutic potential of protease inhibition","volume":"88","author":"Cwilichowska","year":"2022","journal-title":"Mol. Aspects Med."},{"key":"2026082208551690400_cit4","doi-asserted-by":"publisher","first-page":"7341493","DOI":"10.1155\/2022\/7341493","article-title":"Paxlovid:\nMechanism of Action, Synthesis, and In Silico Study","volume":"2022","author":"Marzi","year":"2022","journal-title":"BioMed Res. Int."},{"issue":"22","key":"2026082208551690400_cit5","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.01265-20","article-title":"Development\nof a Fluorescence-Based,\nHigh-Throughput SARS-CoV-2 3CL(pro) Reporter Assay","volume":"94","author":"Froggatt","year":"2020","journal-title":"J. Virol."},{"issue":"11","key":"2026082208551690400_cit6","doi-asserted-by":"publisher","first-page":"4526","DOI":"10.1021\/jacs.8b13042","article-title":"Designing a Green Fluorogenic Protease Reporter by\nFlipping a Beta Strand of GFP for Imaging Apoptosis in Animals","volume":"141","author":"Zhang","year":"2019","journal-title":"J. Am. Chem. Soc."},{"issue":"8","key":"2026082208551690400_cit7","doi-asserted-by":"publisher","first-page":"2212","DOI":"10.1021\/acschembio.2c00364","article-title":"A Modular\nFluorescent Sensor Motif Used to Detect Opioids, Protein-Protein Interactions,\nand Protease Activity","volume":"17","author":"Kroning","year":"2022","journal-title":"ACS Chem. Biol."},{"issue":"4","key":"2026082208551690400_cit8","doi-asserted-by":"publisher","first-page":"555","DOI":"10.1039\/D4CB00276H","article-title":"Functionalization\nof a versatile fluorescent sensor for detecting protease activity\nand temporally gated opioid sensing","volume":"6","author":"Sescil","year":"2025","journal-title":"RSC Chem.\nBiol."},{"issue":"2","key":"2026082208551690400_cit9","doi-asserted-by":"publisher","first-page":"133","DOI":"10.1038\/s41592-020-01010-5","article-title":"Deciphering molecular\ninteractions by proximity labeling","volume":"18","author":"Qin","year":"2021","journal-title":"Nat. Methods"},{"issue":"50","key":"2026082208551690400_cit10","doi-asserted-by":"publisher","first-page":"22933","DOI":"10.1021\/jacs.2c08280","article-title":"Modular Peroxidase-Based\nReporters\nfor Detecting Protease Activity and Protein Interactions with Temporal\nGating","volume":"144","author":"Zhou","year":"2022","journal-title":"J. Am. Chem. Soc."},{"issue":"9","key":"2026082208551690400_cit11","doi-asserted-by":"publisher","first-page":"880","DOI":"10.1038\/nbt.4201","article-title":"Efficient\nproximity labeling in living cells and organisms with TurboID","volume":"36","author":"Branon","year":"2018","journal-title":"Nat. Biotechnol."},{"issue":"22","key":"2026082208551690400_cit12","doi-asserted-by":"publisher","first-page":"12143","DOI":"10.1073\/pnas.1919528117","article-title":"Split-TurboID enables contact-dependent\nproximity labeling in cells","volume":"117","author":"Cho","year":"2020","journal-title":"Proc. Natl. Acad.\nSci. U. S. A."},{"issue":"5","key":"2026082208551690400_cit13","doi-asserted-by":"publisher","first-page":"949","DOI":"10.1016\/S0006-291X(02)00574-0","article-title":"The P1\u2019 specificity\nof tobacco etch virus protease","volume":"294","author":"Kapust","year":"2002","journal-title":"Biochem. Biophys.\nRes. Commun."},{"issue":"11","key":"2026082208551690400_cit14","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0027228","article-title":"Profiling of substrate\nspecificities of 3C-like proteases from group 1, 2a, 2b, and 3 coronaviruses","volume":"6","author":"Chuck","year":"2011","journal-title":"PLoS One"},{"issue":"31","key":"2026082208551690400_cit15","doi-asserted-by":"publisher","first-page":"9340","DOI":"10.1021\/bi963054n","article-title":"Mechanistic\nrole of an NS4A peptide cofactor with the truncated NS3 protease of\nhepatitis C virus: elucidation of the NS4A stimulatory effect via\nkinetic analysis and inhibitor mapping","volume":"36","author":"Landro","year":"1997","journal-title":"Biochemistry"},{"key":"2026082208551690400_cit16","doi-asserted-by":"publisher","first-page":"33","DOI":"10.2147\/HMER.S130103","article-title":"Grazoprevir\/elbasvir\nfor the treatment of adults with\nchronic hepatitis C: a short review on the clinical evidence and place\nin therapy","volume":"10","author":"Sulejmani","year":"2018","journal-title":"Hepatic Med."}],"container-title":["ACS Chemical Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acschembio.6c00334","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/21\/8\/1952\/66116689\/acschembio.6c00334.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acbcct\/article-pdf\/21\/8\/1952\/66116689\/acschembio.6c00334.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,8,22]],"date-time":"2026-08-22T12:55:22Z","timestamp":1787403322000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acbcct\/article\/21\/8\/1952\/5226316\/Biotin-Ligase-Based-Reporter-for-Detecting"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,7,22]]},"references-count":16,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,7,22]]},"published-print":{"date-parts":[[2026,8,21]]}},"URL":"https:\/\/doi.org\/10.1021\/acschembio.6c00334","relation":{},"ISSN":["1554-8929","1554-8937"],"issn-type":[{"value":"1554-8929","type":"print"},{"value":"1554-8937","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,7,22]]}}}