Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP
Sensitive and accurate detection of specific metal ions is important for sensor development and can advance analytical science and support environmental and human medical examinations. Fluorescent proteins (FPs) can be quenched by specific metal ions and spectroscopically show a unique fluorescence-...
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| Format: | Article |
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MDPI AG
2022-05-01
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| Series: | Molecules |
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| Online Access: | https://www.mdpi.com/1420-3049/27/9/2922 |
| _version_ | 1797503647155748864 |
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| author | In Jung Kim Yongbin Xu Ki Hyun Nam |
| author_facet | In Jung Kim Yongbin Xu Ki Hyun Nam |
| author_sort | In Jung Kim |
| collection | DOAJ |
| description | Sensitive and accurate detection of specific metal ions is important for sensor development and can advance analytical science and support environmental and human medical examinations. Fluorescent proteins (FPs) can be quenched by specific metal ions and spectroscopically show a unique fluorescence-quenching sensitivity, suggesting their potential application as FP-based metal biosensors. Since the characteristics of the fluorescence quenching are difficult to predict, spectroscopic analysis of new FPs is important for the development of FP-based biosensors. Here we reported the spectroscopic and structural analysis of metal-induced fluorescence quenching of the photoconvertible fluorescent protein DendFP. The spectroscopic analysis showed that Fe<sup>2+</sup>, Fe<sup>3+</sup>, and Cu<sup>2+</sup> significantly reduced the fluorescence emission of DendFP. The metal titration experiments showed that the dissociation constants (<i>K</i><sub>d</sub>) of Fe<sup>2+</sup>, Fe<sup>3+</sup>, and Cu<sup>2+</sup> for DendFP were 24.59, 41.66, and 137.18 μM, respectively. The tetrameric interface of DendFP, which the metal ions cannot bind to, was analyzed. Structural comparison of the metal-binding sites of DendFP with those of iq-mEmerald and Dronpa suggested that quenchable DendFP has a unique metal-binding site on the β-barrel that does not utilize the histidine pair for metal binding. |
| first_indexed | 2024-03-10T03:53:41Z |
| format | Article |
| id | doaj.art-741170c291b04e0fb09cde9922e92719 |
| institution | Directory Open Access Journal |
| issn | 1420-3049 |
| language | English |
| last_indexed | 2024-03-10T03:53:41Z |
| publishDate | 2022-05-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Molecules |
| spelling | doaj.art-741170c291b04e0fb09cde9922e927192023-11-23T08:51:38ZengMDPI AGMolecules1420-30492022-05-01279292210.3390/molecules27092922Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFPIn Jung Kim0Yongbin Xu1Ki Hyun Nam2Division of Biotechnology, College of Life Sciences and Biotechnology, Korea University, Seoul 02841, KoreaDepartment of Bioengineering, College of Life Science, Dalian Minzu University, Dalian 116600, ChinaDepartment of Life Science, Pohang University of Science and Technology, Pohang 37673, KoreaSensitive and accurate detection of specific metal ions is important for sensor development and can advance analytical science and support environmental and human medical examinations. Fluorescent proteins (FPs) can be quenched by specific metal ions and spectroscopically show a unique fluorescence-quenching sensitivity, suggesting their potential application as FP-based metal biosensors. Since the characteristics of the fluorescence quenching are difficult to predict, spectroscopic analysis of new FPs is important for the development of FP-based biosensors. Here we reported the spectroscopic and structural analysis of metal-induced fluorescence quenching of the photoconvertible fluorescent protein DendFP. The spectroscopic analysis showed that Fe<sup>2+</sup>, Fe<sup>3+</sup>, and Cu<sup>2+</sup> significantly reduced the fluorescence emission of DendFP. The metal titration experiments showed that the dissociation constants (<i>K</i><sub>d</sub>) of Fe<sup>2+</sup>, Fe<sup>3+</sup>, and Cu<sup>2+</sup> for DendFP were 24.59, 41.66, and 137.18 μM, respectively. The tetrameric interface of DendFP, which the metal ions cannot bind to, was analyzed. Structural comparison of the metal-binding sites of DendFP with those of iq-mEmerald and Dronpa suggested that quenchable DendFP has a unique metal-binding site on the β-barrel that does not utilize the histidine pair for metal binding.https://www.mdpi.com/1420-3049/27/9/2922metal biosensorfluorescent proteinDendFPfluorescence quenchingcrystal structure |
| spellingShingle | In Jung Kim Yongbin Xu Ki Hyun Nam Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP Molecules metal biosensor fluorescent protein DendFP fluorescence quenching crystal structure |
| title | Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP |
| title_full | Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP |
| title_fullStr | Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP |
| title_full_unstemmed | Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP |
| title_short | Metal-Induced Fluorescence Quenching of Photoconvertible Fluorescent Protein DendFP |
| title_sort | metal induced fluorescence quenching of photoconvertible fluorescent protein dendfp |
| topic | metal biosensor fluorescent protein DendFP fluorescence quenching crystal structure |
| url | https://www.mdpi.com/1420-3049/27/9/2922 |
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