Design Principles for SuCESsFul Biosensors: Specific Fluorophore/Analyte Binding and Minimization of Fluorophore/Scaffold Interactions
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- 作者：Seymour de Picciotto1 ; Paige M. Dickson2 ; Michael W. Traxlmayr3 ; Bryan S. Marques4 ; Elke Socher5 ; Sixing Zhao1 ; Stephanie Cheung5 ; Jonathan D. Kiefer6 ; A. Joshua Wand4 ; Linda G. Griffith1 ; 4 ; Barbara Imperiali5 ; 7 ; K. Dane Wittrup1 ; 3 ; 8 ; wittrup@MIT.EDU" class="auth_mail" title="E-mail the corresponding author
- 关键词：SuCESsFul ; scaffold conjugated to environment-sensitive fluorophore ; YSD ; yeast surface display ; Fn3 ; 10th type 3 domain of human fibronectin ; FACS ; fluorescence-activated cell sorting ; SUMO ; small ubiquitylation-modified ; DARPin ; designed ankyrin repeat protein ; Sso7d ; small DNA-binding domain derived from Sulfolobus solfataricus ; BTC ; betacellulin ; EGF ; epidermal growth factor ; NBD ; nitrobenz-2-oxa-1 ; 3-diazole ; Badan ; 6-Bromoacetyl-2-dimethylaminonaphthalene ; 4-DMN ; 4-N ; N-dimethylamino-1 ; 8-nap
- 刊名：Journal of Molecular Biology
- 出版年：2016
- 出版时间：9 October 2016
- 年：2016
- 卷：428
- 期：20
- 页码：4228-4241
- 全文大小：1158 K
文摘
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SuCESsFul biosensors report protein interactions via environment-sensitive dyes.
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We observed that the current design principles are insufficient.
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Fluorophore-binding pockets are required for strong fluorescence activation.
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Scaffold/fluorophore interaction can raise background fluorescence.
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Using our new design criteria, we engineered the most responsive sensor to date.