Development of a Novel Blue Fluorescent Gene-encoded Calcium Indicator Modified from GCaMP3

Yum, Chorok; Ahn, Taekyoung; Shim, Won‐Sik

doi:10.1007/s10895-020-02606-y

Cited by 1 publication

(2 citation statements)

References 20 publications

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“…Still, this structure solved at 1.8 Å resolution revealed a lack of electron density for the phenolic moiety of the chromophore. We, however, were able to trace one conformation of this group with partial occupancy, which is somewhat similar to those found in the blue FP-mTagBFP [14]. A detailed analysis of the chromophore environment and its comparison with two related structurally characterized blue FPs-Blue102 and mTagBFP-revealed a number of characteristic differences.…”

Section: Introductionsupporting

confidence: 64%

“…A number of blue FPs (BFP) have been described to date [14,[17][18][19] as well as fluorescent timers that change their color from blue to red upon maturation [1][2][3]5]. A monomeric fluorescent timer, Fast-FT, made from mCherry FP, was previously structurally characterized together with its blue variant (Blue102, PDB ID-3LF4) with blocked blue-to-red conversion [5].…”

Section: Comparison With Blue102-a Blue Form Of Fluorescent Timer Fas...mentioning

confidence: 99%

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Combined Structural and Computational Study of the mRubyFT Fluorescent Timer Locked in Its Blue Form

Boyko

Khrenova

Дороватовский

et al. 2023

IJMS

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The mRubyFT is a monomeric genetically encoded fluorescent timer based on the mRuby2 fluorescent protein, which is characterized by the complete maturation of the blue form with the subsequent conversion to the red one. It has higher brightness in mammalian cells and higher photostability compared with other fluorescent timers. A high-resolution structure is a known characteristic of the mRubyFT with the red form chromophore, but structural details of its blue form remain obscure. In order to obtain insight into this, we obtained an S148I variant of the mRubyFT (mRubyFTS148I) with the blocked over time blue form of the chromophore. X-ray data at a 1.8 Å resolution allowed us to propose a chromophore conformation and its interactions with the neighboring residues. The imidazolidinone moiety of the chromophore is completely matured, being a conjugated π-system. The methine bridge is not oxidized in the blue form bringing flexibility to the phenolic moiety that manifests itself in poor electron density. Integration of these data with the results of molecular dynamic simulation disclosed that the OH group of the phenolic moiety forms a hydrogen bond with the side chain of the T163 residue. A detailed comparison of mRubyFTS148I with other available structures of the blue form of fluorescent proteins, Blue102 and mTagBFP, revealed a number of characteristic differences. Molecular dynamic simulations with the combined quantum mechanic/molecular mechanic potentials demonstrated that the blue form exists in two protonation states, anion and zwitterion, both sharing enolate tautomeric forms of the C=C–O− fragment. These two forms have similar excitation energies, as evaluated by calculations. Finally, excited state molecular dynamic simulations showed that excitation of the chromophore in both protonation states leads to the same anionic fluorescent state. The data obtained shed light on the structural features and spectral properties of the blue form of the mRubyFT timer.

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Section: Introductionsupporting

confidence: 64%

Section: Comparison With Blue102-a Blue Form Of Fluorescent Timer Fas...mentioning

confidence: 99%

Combined Structural and Computational Study of the mRubyFT Fluorescent Timer Locked in Its Blue Form

Boyko

Khrenova

Дороватовский

et al. 2023

IJMS

View full text Add to dashboard Cite

show abstract

Development of a Novel Blue Fluorescent Gene-encoded Calcium Indicator Modified from GCaMP3

Cited by 1 publication

References 20 publications

Combined Structural and Computational Study of the mRubyFT Fluorescent Timer Locked in Its Blue Form

Combined Structural and Computational Study of the mRubyFT Fluorescent Timer Locked in Its Blue Form

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