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
Human and Animal Cells HeLa
DNA material Gamillus/pcDNA3 (RDB19864)