Design of small monomeric and highly bright near-infrared fluorescent proteins
| Autor: | Kai-Hong Zhao, Wen-Long Ding, Chao-Di Kong, Xian-Dan Li, Zi-Zhu Tan, Bao-Qing Zhao, Ya-Nan Hou |
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| Rok vydání: | 2019 |
| Předmět: |
Models
Molecular Brightness Light Infrared Rays Protein Conformation Photochemistry Cyanobacteria Fluorescence Absorbance chemistry.chemical_compound Phycocyanobilin Bacterial Proteins Phycobilins Escherichia coli Animals Humans Molecular Biology Biliverdin Near-infrared spectroscopy Biliverdine Optical Imaging Phycocyanin Cell Biology Chromophore Luminescent Proteins Monomer HEK293 Cells chemistry Microscopy Fluorescence HeLa Cells |
| Zdroj: | Biochimica et biophysica acta. Molecular cell research. 1866(10) |
| ISSN: | 1879-2596 |
| Popis: | Due to the low absorbance in the far-red (FR) and near-infrared (NIR) “optical window”, NIR fluorescent proteins (FPs) are powerful tools for deep imaging. Here, we report three new, highly bright NIR FPs termed BDFP1.8, BDFP1.8:1.8 (tandem BDFP1.8) and BDFP1.9, which evolved from a previously reported FR FP, BDFP1.6: a derivative of ApcF2 from Chroococcidiopsis thermalis sp. PCC7203. ApcF2 binds phycocyanobilin (PCB) non-covalently, while BDFPs, the derivatives of ApcF2, can bind biliverdin (BV) covalently. We identified that dimeric BDFP1.8 and monomeric BDFP1.8:1.8 have a 2.4-and 4.4-fold higher effective brightness, respectively, than iRFP720, which has the highest effective brightness among the reported NIR FPs. Monomeric DBFP1.9 (17 kDa) has one of the smallest masses among highly bright FPs in the FR and NIR regions. Enhancing the affinity between the apo-proteins and the BV chromophore is an effective method to improve the effective brightness of biliprotein FPs. Moreover, BDFP1.8 and 1.9 exhibit higher stability to temperature, pH and light than iRFP720. Finally, the highly bright NIR BDFP1.8 together with FR BDFP1.6 could effectively biolabel cells in dual colors. |
| Databáze: | OpenAIRE |
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