7Z7Q

Structure of the T207D single-point mutant of the fluorescent protein NeonCyan1 at pH 6.5

Summary for 7Z7Q

• Entry DOI: 10.2210/pdb7z7q/pdb
• Descriptor: NeonCyan1 (2 entities in total)
• Functional Keywords: mneongreen, trp-based fluorescent chromophore, rational design, fluorescent protein
• Biological source: Branchiostoma lanceolatum (amphioxus)
• Total number of polymer chains: 4
• Total formula weight: 106626.94

Authors

Duarte, K.,Dupuy, J.,Royant, A. (deposition date: 2022-03-16, release date: 2022-05-04, Last modification date: 2024-10-16)

Primary citation

Zarowny, L.,Clavel, D.,Johannson, R.,Duarte, K.,Depernet, H.,Dupuy, J.,Baker, H.,Brown, A.,Royant, A.,Campbell, R.E.
Cyan fluorescent proteins derived from mNeonGreen.
Protein Eng.Des.Sel., 35:-, 2022

PubMed Abstract: mNeonGreen, an engineered green fluorescent protein (GFP) derived from lancelet, is one of the most brightly fluorescent homologs of Aequorea victoria jellyfish GFP (avGFP) yet reported. In this work, we investigated whether this bright fluorescence might be retained in homologs of mNeonGreen with modified chromophore structures and altered fluorescent hues. We found mNeonGreen to be generally less tolerant than avGFP to chromophore modification by substitution of the key chromophore-forming tyrosine residue with other aromatic amino acids. However, we were ultimately successful in creating a variant, designated as NeonCyan1, with a tryptophan-derived cyan fluorescent protein (CFP)-type chromophore, and two additional mutants with distinct spectral hues. Structural, computational, and photophysical characterization of NeonCyan1 and its variants provided insight into the factors that control the fluorescence emission color. Though not recommended as replacements for contemporary CFP variants, we demonstrate that NeonCyan1 variants are potentially suitable for live cell imaging applications.

PubMed: 35417013
DOI: 10.1093/protein/gzac004

Experimental method

X-RAY DIFFRACTION (1.6 Å)