Collagen-Immobilized Extracellular FRET Reporter for Visualizing Protease Activity Secreted by Living Cells
DC Field | Value | Language |
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dc.contributor.author | Lee, Hawon | - |
dc.contributor.author | Kim, Se-jeong | - |
dc.contributor.author | Shin, Heungsoo | - |
dc.contributor.author | Kim, Young-Pil | - |
dc.date.accessioned | 2021-08-02T09:52:33Z | - |
dc.date.available | 2021-08-02T09:52:33Z | - |
dc.date.created | 2021-05-12 | - |
dc.date.issued | 2020-03 | - |
dc.identifier.issn | 2379-3694 | - |
dc.identifier.uri | https://scholarworks.bwise.kr/hanyang/handle/2021.sw.hanyang/10672 | - |
dc.description.abstract | Despite the diverse roles of cell-secreted proteases in the extracellular matrix (ECM), classical methods to analyze protease activity have not been explored at the cell culture site. Here, we report a stable, matrix-sticky, and protease-sensitive extracellular reporter that comprises a collagen-binding protein and a Forster resonance energy transfer (FRET) coupler of an enhanced green fluorescent protein and a small dye molecule. The extracellular FRET reporter via split intein-mediated protein trans-splicing is able to adhere to collagen matrices, leading to fluorescence changes by matrix metalloproteinase-2 (MMP2) activity during living cell culture without impeding cell viability. When a proMMP2 mutant (Y581A) with altered protease secretion and activity was transfected into cancer cells, the reporter revealed a dramatic reduction in MMP2 activity in both two- and three-dimensional culture systems, compared with cells transfected with wild-type proMMP2. Our reporter is immediately amenable to monitor protease activity in diverse ECM-resident cells as well as to study protease-related extracellular signaling and tissue remodeling. | - |
dc.language | 영어 | - |
dc.language.iso | en | - |
dc.publisher | AMER CHEMICAL SOC | - |
dc.title | Collagen-Immobilized Extracellular FRET Reporter for Visualizing Protease Activity Secreted by Living Cells | - |
dc.type | Article | - |
dc.contributor.affiliatedAuthor | Kim, Young-Pil | - |
dc.identifier.doi | 10.1021/acssensors.9b01456 | - |
dc.identifier.scopusid | 2-s2.0-85080139437 | - |
dc.identifier.wosid | 000526377600007 | - |
dc.identifier.bibliographicCitation | ACS SENSORS, v.5, no.3, pp.655 - 664 | - |
dc.relation.isPartOf | ACS SENSORS | - |
dc.citation.title | ACS SENSORS | - |
dc.citation.volume | 5 | - |
dc.citation.number | 3 | - |
dc.citation.startPage | 655 | - |
dc.citation.endPage | 664 | - |
dc.type.rims | ART | - |
dc.type.docType | Article | - |
dc.description.journalClass | 1 | - |
dc.description.isOpenAccess | N | - |
dc.description.journalRegisteredClass | scie | - |
dc.description.journalRegisteredClass | scopus | - |
dc.relation.journalResearchArea | Chemistry | - |
dc.relation.journalResearchArea | Science & Technology - Other Topics | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Multidisciplinary | - |
dc.relation.journalWebOfScienceCategory | Chemistry, Analytical | - |
dc.relation.journalWebOfScienceCategory | Nanoscience & Nanotechnology | - |
dc.subject.keywordPlus | Cell culture | - |
dc.subject.keywordPlus | Cells | - |
dc.subject.keywordPlus | Collagen | - |
dc.subject.keywordPlus | Energy transfer | - |
dc.subject.keywordPlus | Forster resonance energy transfer | - |
dc.subject.keywordPlus | Military electronic countermeasures | - |
dc.subject.keywordPlus | 5-carboxytetramethylrhodamine succinimidyl ester | - |
dc.subject.keywordPlus | collagen type 1 | - |
dc.subject.keywordPlus | coloring agent | - |
dc.subject.keywordPlus | enhanced green fluorescent protein | - |
dc.subject.keywordPlus | gelatinase A | - |
dc.subject.keywordPlus | green fluorescent protein | - |
dc.subject.keywordPlus | recombinant protein | - |
dc.subject.keywordPlus | rhodamine | - |
dc.subject.keywordPlus | Enhanced green fluorescent protein | - |
dc.subject.keywordPlus | Extracellular matrices | - |
dc.subject.keywordPlus | FRET | - |
dc.subject.keywordPlus | intein | - |
dc.subject.keywordPlus | Matrix metalloproteinase-2 | - |
dc.subject.keywordPlus | Protease activities | - |
dc.subject.keywordPlus | Resonance energy transfer | - |
dc.subject.keywordPlus | Three-dimensional culture system | - |
dc.subject.keywordPlus | chemistry | - |
dc.subject.keywordPlus | confocal microscopy | - |
dc.subject.keywordPlus | extracellular matrix | - |
dc.subject.keywordPlus | fluorescence resonance energy transfer | - |
dc.subject.keywordPlus | genetics | - |
dc.subject.keywordPlus | human | - |
dc.subject.keywordPlus | metabolism | - |
dc.subject.keywordPlus | protein splicing | - |
dc.subject.keywordPlus | tumor cell line | - |
dc.subject.keywordPlus | Cell signaling | - |
dc.identifier.url | https://pubs.acs.org/doi/10.1021/acssensors.9b01456 | - |
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