Treffer: StayRose: A photostable StayGold derivative redshifted by genetic code expansion.

Title:
StayRose: A photostable StayGold derivative redshifted by genetic code expansion.
Authors:
Scott W; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK; Department of Chemistry, University of Warwick, Coventry, UK., Ivorra-Molla E; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK., Akhuli D; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK., Massam-Wu T; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK., Lysyganicz PK; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK., Walsh R; Josephine Bay Paul Center, Marine Biological Laboratory, Woods Hole, Massachusetts, USA., Parent M; Josephine Bay Paul Center, Marine Biological Laboratory, Woods Hole, Massachusetts, USA., Cook J; School of Life Sciences, University of Warwick, Coventry, UK., Song L; Department of Chemistry, University of Warwick, Coventry, UK., Kumar A; Josephine Bay Paul Center, Marine Biological Laboratory, Woods Hole, Massachusetts, USA., Schneider F; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK., Mishima M; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK. Electronic address: M.Mishima@warwick.ac.uk., Crow A; School of Life Sciences, University of Warwick, Coventry, UK. Electronic address: Allister.Crow@warwick.ac.uk., Balasubramanian MK; Centre for Mechanochemical Cell Biology and Division of Biomedical Sciences, Warwick Medical School, University of Warwick, Coventry, UK. Electronic address: M.K.Balasubramanian@warwick.ac.uk.
Source:
The Journal of biological chemistry [J Biol Chem] 2025 Dec; Vol. 301 (12), pp. 110832. Date of Electronic Publication: 2025 Oct 16.
Publication Type:
Journal Article
Language:
English
Journal Info:
Publisher: Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology Country of Publication: United States NLM ID: 2985121R Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1083-351X (Electronic) Linking ISSN: 00219258 NLM ISO Abbreviation: J Biol Chem Subsets: MEDLINE
Imprint Name(s):
Publication: 2021- : [New York, NY] : Elsevier Inc. on behalf of American Society for Biochemistry and Molecular Biology
Original Publication: Baltimore, MD : American Society for Biochemistry and Molecular Biology
MeSH Terms:
Luminescent Proteins*/genetics , Luminescent Proteins*/chemistry , Luminescent Proteins*/metabolism , Genetic Code* , Green Fluorescent Proteins*/genetics , Green Fluorescent Proteins*/chemistry , Green Fluorescent Proteins*/metabolism, Zebrafish/embryology ; Animals ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Red Fluorescent Protein ; Tyrosine/chemistry ; Tyrosine/analogs & derivatives ; Tyrosine/genetics ; Crystallography, X-Ray
Contributed Indexing:
Keywords: 3-aminotyrosine; StayGold; StayRose; crystal structure; fluorescence; genetic code expansion; mCherry; microscopy; protein engineering; synthetic biology
Substance Nomenclature:
0 (Red Fluorescent Protein)
0 (Luminescent Proteins)
147336-22-9 (Green Fluorescent Proteins)
42HK56048U (Tyrosine)
Entry Date(s):
Date Created: 20251018 Date Completed: 20251230 Latest Revision: 20260106
Update Code:
20260106
PubMed Central ID:
PMC12661434
DOI:
10.1016/j.jbc.2025.110832
PMID:
41109344
Database:
MEDLINE

Weitere Informationen

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.
(Copyright © 2025 The Authors. Published by Elsevier Inc. All rights reserved.)

Conflict of interest The authors declare that they have no conflicts of interest with the contents of this article.