Development of a Protease Biosensor Based on a Dimerization-Dependent Red Fluorescent Protein Article Swipe
Aaron C. Mitchell
,
Spencer C. Alford
,
Sean A. Yamada‐Hunter
,
Deepti Kannan
,
R. Andres Parra Sperberg
,
Cheryl Chang
,
Jennifer R. Cochran
·
YOU?
·
· 2017
· Open Access
·
· DOI: https://doi.org/10.1021/acschembio.7b00715
YOU?
·
· 2017
· Open Access
·
· DOI: https://doi.org/10.1021/acschembio.7b00715
Dysregulated activity of the protease matriptase is a key contributor to aggressive tumor growth, cancer metastasis, and osteoarthritis. Methods for the detection and quantification of matriptase activity and inhibition would be useful tools. To address this need, we developed a matriptase-sensitive protein biosensor based on a dimerization-dependent red fluorescent protein (ddRFP) reporter system. In this platform, two adjoining protein domains, connected by a protease-labile linker, produce fluorescence when assembled and are nonfluorescent when the linker is cleaved by matriptase. A panel of ddRFP-based matriptase biosensor designs was created that contained different linker lengths between the protein domains. These constructs were characterized for linker-specific cleavage, matriptase activity, and matriptase selectivity; a biosensor containing a RSKLRVGGH linker (termed B4) was expressed at high yields and displayed both high catalytic efficiency and matriptase specificity. This biosensor detects matriptase inhibition by soluble and yeast cell surface expressed inhibitor domains with up to a 5-fold dynamic range and also detects matriptase activity expressed by human cancer cell lines. In addition to matriptase, we highlight a strategy that can be used to create effective biosensors for quantifying activity and inhibition of other proteases of interest.
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Metadata
- Type
- article
- Language
- en
- Landing Page
- https://doi.org/10.1021/acschembio.7b00715
- OA Status
- green
- Cited By
- 20
- References
- 34
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W2767964081
All OpenAlex metadata
Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W2767964081Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.1021/acschembio.7b00715Digital Object Identifier
- Title
-
Development of a Protease Biosensor Based on a Dimerization-Dependent Red Fluorescent ProteinWork title
- Type
-
articleOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2017Year of publication
- Publication date
-
2017-11-10Full publication date if available
- Authors
-
Aaron C. Mitchell, Spencer C. Alford, Sean A. Yamada‐Hunter, Deepti Kannan, R. Andres Parra Sperberg, Cheryl Chang, Jennifer R. CochranList of authors in order
- Landing page
-
https://doi.org/10.1021/acschembio.7b00715Publisher landing page
- Open access
-
YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
-
https://www.ncbi.nlm.nih.gov/pmc/articles/6453536Direct OA link when available
- Concepts
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Linker, Biosensor, Proteases, Protease, Chemistry, Cleavage (geology), Biochemistry, Fluorescence, Cancer cell, Cell biology, Biology, Enzyme, Cancer, Computer science, Fracture (geology), Paleontology, Physics, Operating system, Quantum mechanics, GeneticsTop concepts (fields/topics) attached by OpenAlex
- Cited by
-
20Total citation count in OpenAlex
- Citations by year (recent)
-
2025: 1, 2024: 3, 2023: 2, 2022: 4, 2021: 5Per-year citation counts (last 5 years)
- References (count)
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34Number of works referenced by this work
- Related works (count)
-
10Other works algorithmically related by OpenAlex
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| abstract_inverted_index.activity | 1, 26, 156, 181 |
| abstract_inverted_index.addition | 164 |
| abstract_inverted_index.domains, | 59 |
| abstract_inverted_index.domains. | 96 |
| abstract_inverted_index.protease | 4 |
| abstract_inverted_index.reporter | 51 |
| abstract_inverted_index.strategy | 170 |
| abstract_inverted_index.RSKLRVGGH | 113 |
| abstract_inverted_index.activity, | 105 |
| abstract_inverted_index.adjoining | 57 |
| abstract_inverted_index.assembled | 68 |
| abstract_inverted_index.biosensor | 42, 84, 110, 132 |
| abstract_inverted_index.catalytic | 126 |
| abstract_inverted_index.cleavage, | 103 |
| abstract_inverted_index.connected | 60 |
| abstract_inverted_index.contained | 89 |
| abstract_inverted_index.detection | 21 |
| abstract_inverted_index.developed | 38 |
| abstract_inverted_index.different | 90 |
| abstract_inverted_index.displayed | 123 |
| abstract_inverted_index.effective | 177 |
| abstract_inverted_index.expressed | 118, 142, 157 |
| abstract_inverted_index.highlight | 168 |
| abstract_inverted_index.inhibitor | 143 |
| abstract_inverted_index.interest. | 188 |
| abstract_inverted_index.platform, | 55 |
| abstract_inverted_index.proteases | 186 |
| abstract_inverted_index.aggressive | 11 |
| abstract_inverted_index.biosensors | 178 |
| abstract_inverted_index.constructs | 98 |
| abstract_inverted_index.containing | 111 |
| abstract_inverted_index.efficiency | 127 |
| abstract_inverted_index.inhibition | 28, 135, 183 |
| abstract_inverted_index.matriptase | 5, 25, 83, 104, 107, 129, 134, 155 |
| abstract_inverted_index.contributor | 9 |
| abstract_inverted_index.ddRFP-based | 82 |
| abstract_inverted_index.fluorescent | 48 |
| abstract_inverted_index.matriptase, | 166 |
| abstract_inverted_index.matriptase. | 78 |
| abstract_inverted_index.metastasis, | 15 |
| abstract_inverted_index.quantifying | 180 |
| abstract_inverted_index.Dysregulated | 0 |
| abstract_inverted_index.fluorescence | 66 |
| abstract_inverted_index.selectivity; | 108 |
| abstract_inverted_index.specificity. | 130 |
| abstract_inverted_index.characterized | 100 |
| abstract_inverted_index.nonfluorescent | 71 |
| abstract_inverted_index.quantification | 23 |
| abstract_inverted_index.linker-specific | 102 |
| abstract_inverted_index.osteoarthritis. | 17 |
| abstract_inverted_index.protease-labile | 63 |
| abstract_inverted_index.matriptase-sensitive | 40 |
| abstract_inverted_index.dimerization-dependent | 46 |
| cited_by_percentile_year.max | 98 |
| cited_by_percentile_year.min | 90 |
| countries_distinct_count | 1 |
| institutions_distinct_count | 7 |
| sustainable_development_goals[0].id | https://metadata.un.org/sdg/3 |
| sustainable_development_goals[0].score | 0.7400000095367432 |
| sustainable_development_goals[0].display_name | Good health and well-being |
| citation_normalized_percentile.value | 0.77606885 |
| citation_normalized_percentile.is_in_top_1_percent | False |
| citation_normalized_percentile.is_in_top_10_percent | False |