Jennifer Sescil, Isabel Solowiej, Guanwei Zhou, Tae Heo, Emma Fu, Wenjing Wang
{"title":"Biotin Ligase-Based Reporter for Detecting Protease Activity","authors":"Jennifer Sescil, Isabel Solowiej, Guanwei Zhou, Tae Heo, Emma Fu, Wenjing Wang","doi":"10.1021/acschembio.6c00334","DOIUrl":null,"url":null,"abstract":"<p>Protease cleavage is a crucial process, associated with\r\ncellular\r\nsignaling and viral infection. Here, we present a novel sensing motif\r\nbased on a proximity-dependent biotin ligase, TurboID. The sensor,\r\nBiotin Annotating Genetically Encoded Ligase-based Sensor (BAGELS),\r\nwas created by engineering TurboID such that it is only active following\r\nprotease activity, linking cleavage events to the biotinylation of\r\nproximal proteins. BAGELS produces a signal-to-background ratio of\r\napproximately 48 when tested with and without protease. This reporter\r\nrepresents the first protease activity sensor with biotin ligase readout\r\nin neurons and HEK293T cells. We demonstrate the reporter’s\r\nuse for characterization of protease inhibitors and multiplexed imaging\r\nwith fluorogenic sensors. Overall, this sensor offers a versatile,\r\nenzymatically amplified biotin-based readout compatible with biotin-based\r\ndetection techniques.</p>","PeriodicalId":11,"journal":{"name":"ACS Chemical Biology","volume":"21 8","pages":"1952–1957"},"PeriodicalIF":3.6000,"publicationDate":"2026-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"ACS Chemical Biology","FirstCategoryId":"99","ListUrlMain":"https://pubs.acs.org/doi/10.1021/acschembio.6c00334","RegionNum":2,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"2026/7/22 0:00:00","PubModel":"Epub","JCR":"Q2","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
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’s
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.
期刊介绍：
ACS Chemical Biology provides an international forum for the rapid communication of research that broadly embraces the interface between chemistry and biology.
The journal also serves as a forum to facilitate the communication between biologists and chemists that will translate into new research opportunities and discoveries. Results will be published in which molecular reasoning has been used to probe questions through in vitro investigations, cell biological methods, or organismic studies.
We welcome mechanistic studies on proteins, nucleic acids, sugars, lipids, and nonbiological polymers. The journal serves a large scientific community, exploring cellular function from both chemical and biological perspectives. It is understood that submitted work is based upon original results and has not been published previously.
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
