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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Chen, Lingcheng Wang, Chao Song, Xinbo Xiao, Yi |
| Description | Country affiliation: China Author Affiliation: Wang C ( State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China.); Song X ( State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China.); Chen L ( State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China.); Xiao Y ( State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian 116024, China. Electronic address: xiaoyi@dlut.edu.cn.) |
| Abstract | Viscosity, as one of the major factors of intracellular microenvironment, influences the function of proteins. To detect local micro-viscosity of a protein, it is a precondition to apply a viscosity sensor for specifically target to proteins. However, all the reported small-molecule probes are just suitable for sensing/imaging of macro-viscosity in biological fluids of entire cells or organelles. To this end, we developed a hybrid sensor BDP-V BG by connecting a viscosity-sensitive boron-dipyrromethene (BODIPY) molecular rotor (BDP-V) to O -benzylguanine (BG) for specific detection of local micro-viscosity of SNAP-tag fused proteins. We measured and calculated the reaction efficiency between the sensor and SNAP-tag protein in vitro to confirm the high labeling specificity. We also found that the labeling reaction results in a 53-fold fluorescence enhancement for the rotor, which qualifies it as a wash-free sensor with ignorable background fluorescence. The high sensitivity of protein labeled sensor (BDP-V-SNAP) to the changes of local viscosity was evaluated by detecting the enhancement of fluorescence lifetimes. Further, with the sensor BDP-V BG, we achieved high specific labeling of cells expressing two SNAP-tag fused proteins (nuclear histone H2B and mitochondrial COX8A). Two-photon excited fluorescence lifetime imaging revealed that, the micro-viscosities nearby the SNAP-tag fused two proteins are distinct. The different changes of local micro-viscosity of SNAP-tag fused histone protein in apoptosis induced by three nucleus-targeted drugs were also characterized for the first time. |
| ISSN | 09565663 |
| Issue Number | Pt 2 |
| Journal | Biosensors and Bioelectronics |
| Volume Number | 89 |
| e-ISSN | 18734235 |
| Language | English |
| Publisher | Elsevier |
| Publisher Date | 2016-11-10 |
| Publisher Place | Great Britain (UK) |
| Access Restriction | One Nation One Subscription (ONOS) |
| Subject Keyword | Discipline Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Nanoscience and Nanotechnology Medicine Biophysics Biomedical Engineering Biotechnology Electrochemistry |
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