Reference - Detail
| RRC ID | 58007 |
|---|---|
| Author | Shinoda H, Lu K, Nakashima R, Wazawa T, Noguchi K, Matsuda T, Nagai T. |
| Title | Acid-Tolerant Reversibly Switchable Green Fluorescent Protein for Super-resolution Imaging under Acidic Conditions. |
| Journal | Cell Chem Biol |
| Abstract |
Reversibly switchable fluorescent proteins (RSFPs) are crucial tags for super-resolution observation of protein localization and dynamics inside living cells. However, due to the high fluorescence pKa (∼5-6) of most RSFPs, their usage in acidic conditions (pH 4.5-6.0) has been limited. Here, we investigated a new photochromic mechanism in Gamillus, a recently developed green fluorescent protein with acid tolerance. Gamillus exhibits negative switching with especially high contrast in acidic conditions, and its off switching is caused by trans-to-cis isomerization of the chromophore hydroxyphenyl ring that accompanies protonation. Through a combination of rational design and saturation mutagenesis, we developed two variants with enhanced switching contrasts and off-switching speeds, designated rsGamillus-S and rsGamillus-F, respectively. The fluorescence intensity, off-switching speed, and switching contrast of the rsGamillus variants are only slightly affected by changes in pH between 4.5 and 7.5. Exploiting these properties, we succeeded in high-contrast super-resolution imaging of cellular architectures in acidic conditions. |
| Volume | 26(10) |
| Pages | 1469-1479.e6 |
| Published | 2019-10-17 |
| DOI | 10.1016/j.chembiol.2019.07.012 |
| PII | S2451-9456(19)30243-0 |
| PMID | 31422907 |
| MeSH | Crystallography, X-Ray Green Fluorescent Proteins / chemistry* HeLa Cells Humans Hydrogen-Ion Concentration Models, Molecular Photochemical Processes Tumor Cells, Cultured |
| IF | 6.762 |
| Times Cited | 1 |
| Altmetric score |
オルトメトリクス指標項目
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| The most frequently cited source | X(Twitter) |
| Total number of mentions | 56 |
| Altmetric score changes over past 6months | 0.0 |
| Resource | |
| Human and Animal Cells | COS-7(RCB0539) HeLa |
| DNA material | rsGamillus-S/pcDNA3 (RDB19867) rsGamillus-F/pcDNA3 (RDB19868) |