Saved in:
| Main Authors: | , , , , , , , , , , , , , |
|---|---|
| Format: | Artículo científico |
| Language: | en |
| Published: |
The Journal of biological chemistry
2025
|
| Subjects: | |
| Online Access: | https://pubmed.ncbi.nlm.nih.gov/41109344/ |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
Table of Contents:
- StayRose: A photostable StayGold derivative redshifted by genetic code expansion. Scott, Will Ivorra-Molla, Esther Akhuli, Dipayan Massam-Wu, Teresa Lysyganicz, Pawel K Walsh, Rylie Parent, Matthew Cook, Jonathan Song, Lijiang Kumar, Abhishek Schneider, Falk Mishima, Masanori Crow, Allister Balasubramanian, Mohan K Zebrafish Animals Escherichia coli Red Fluorescent Protein Luminescent Proteins Genetic Code Green Fluorescent Proteins Tyrosine Crystallography, X-Ray Photobleaching of fluorescent proteins often limits the acquisition of high-quality images in microscopy. StayGold, a novel dimeric GFP recently monomerized through sequence engineering, addresses this challenge with its high photostability. There is now a focus on producing different colored StayGold derivatives to facilitate concurrent tagging of multiple targets. The unnatural amino acid 3-aminotyrosine has previously been shown to redshift superfolder GFP upon incorporation into its chromophore via genetic code expansion. Here, we apply the same strategy to redshift StayGold through substitution of tyrosine-58 with 3-aminotyrosine. The resultant red fluorescent protein, StayRose, shows an excitation wavelength maximum of 530 nm and an emission wavelength maximum of 588 nm. Importantly, the monomeric mStayRose retains the favorable photostability in vivo in Escherichia coli and zebrafish embryos. A high-resolution crystal structure of StayRose confirms the modified structure of the amino chromophore within an unperturbed 3D fold. Although reliant on genetic code expansion, StayRose provides an important step toward developing redshifted StayGold derivatives.