6J1A
Photoswitchable fluorescent protein Gamillus, off-state
Summary for 6J1A
| Entry DOI | 10.2210/pdb6j1a/pdb |
| Descriptor | Green fluorescent protein, CHLORIDE ION, GLYCEROL, ... (5 entities in total) |
| Functional Keywords | acid-tolerant, photoswitchable, cis-chromophore, fluorescent protein |
| Biological source | Olindias |
| Total number of polymer chains | 1 |
| Total formula weight | 30795.75 |
| Authors | Nakashima, R.,Sakurai, K.,shinoda, H.,Matsuda, T.,Nagai, T. (deposition date: 2018-12-28, release date: 2019-11-06, Last modification date: 2023-11-22) |
| Primary citation | Shinoda, H.,Lu, K.,Nakashima, R.,Wazawa, T.,Noguchi, K.,Matsuda, T.,Nagai, T. Acid-Tolerant Reversibly Switchable Green Fluorescent Protein for Super-resolution Imaging under Acidic Conditions. Cell Chem Biol, 26:1469-1479.e6, 2019 Cited by PubMed 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 pK (∼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. PubMed: 31422907DOI: 10.1016/j.chembiol.2019.07.012 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.96 Å) |
Structure validation
Download full validation report
