| RRC ID |
67313
|
| Author |
Shinoda H, Ma Y, Nakashima R, Sakurai K, Matsuda T, Nagai T.
|
| Title |
Acid-Tolerant Monomeric GFP from Olindias formosa.
|
| Journal |
Cell Chem Biol
|
| Abstract |
The fluorescent protein (FP) color palette has greatly contributed to the visualization of molecular and cellular processes. However, most FPs lose fluorescence at a pH lower than their neutral pKa (∼6), and this has hampered their application in acidic organelles (pH ∼4.5-6.0). Currently, several cyan- and red-colored acid-tolerant FPs are available; however, there are few reports of acid-tolerant green FPs (GFPs) that are practically applicable to bioimaging. Here, we developed the acid-tolerant monomeric GFP "Gamillus" from the jellyfish Olindias formosa, with excellent brightness, maturation speed, and photostability. Results from X-ray crystallography and point mutagenesis suggest that across a broad pH range the acid tolerance is attributed to stabilization of deprotonation in the chromophore phenyl ring by forming a unique trans configuration. We demonstrate that Gamillus can serve as a molecular tag suitable for imaging in acidic organelles through autophagy-mediated molecular tracking to lysosomes.
|
| Volume |
25(3)
|
| Pages |
330-338.e7
|
| Published |
2018-3-15
|
| DOI |
10.1016/j.chembiol.2017.12.005
|
| PII |
S2451-9456(17)30432-4
|
| PMID |
29290624
|
| MeSH |
Animals
Crystallography, X-Ray
Green Fluorescent Proteins / chemistry
Green Fluorescent Proteins / genetics
Green Fluorescent Proteins / metabolism*
Hydrogen-Ion Concentration
Hydrozoa / metabolism*
Lysosomes / metabolism
Protein Structure, Tertiary
Recombinant Proteins / biosynthesis
Recombinant Proteins / chemistry
Recombinant Proteins / isolation & purification
Solubility
Spectrometry, Fluorescence
|
| IF |
6.762
|
| Resource |
| Human and Animal Cells |
HeLa |
| DNA material |
Gamillus/pcDNA3 (RDB19864) |
