Recombinant light-sensitive photoprotein berovin from ctenophore Beroe abyssicola: Bioluminescence and absorbance characteristics

Burakova, Ludmila P.; Колмакова, А. А.; Vysotski, Eugene S.

doi:10.1016/j.bbrc.2022.07.079

Cited by 3 publications

(1 citation statement)

References 23 publications

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“…Since CTM-v is identified as the main product of obelin-v bioluminescence by structural data and can be considered as a product of a "dark" pathway of the decomposition of dioxetanone intermediate, it explains the low bioluminescence activity of obelin-v relative to that of obelin activated with unmodified CTZ. This explanation is also supported by the quantum yields of bioluminescence reaction (Φ BL ) of cnidarian and ctenophore Ca 2+ -regulated photoproteins 34,35 . The Φ BL value of aequorin bioluminescence amounts to 0.19, while Φ BL of berovin is more than twice lower (0.083).…”

Section: Discussionmentioning

confidence: 74%

Crystal structure of semi-synthetic obelin-v after calcium induced bioluminescence implies coelenteramine as the main reaction product

Natashin

Eremeeva

Shevtsov

et al. 2022

Sci Rep

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Coelenterazine-v (CTZ-v), a synthetic vinylene-bridged π-extended derivative, is able to significantly alter bioluminescence spectra of different CTZ-dependent luciferases and photoproteins by shifting them towards longer wavelengths. However, Ca2+-regulated photoproteins activated with CTZ-v display very low bioluminescence activities that hampers its usage as a substrate of photoprotein bioluminescence. Here, we report the crystal structure of semi-synthetic Ca2+-discharged obelin-v bound with the reaction product determined at 2.1 Å resolution. Comparison of the crystal structure of Ca2+-discharged obelin-v with those of other obelins before and after bioluminescence reaction reveals no considerable changes in the overall structure. However, the drastic changes in CTZ-binding cavity are observed owing to the completely different reaction product, coelenteramine-v (CTM-v). Since CTM-v is certainly the main product of obelin-v bioluminescence and is considered to be a product of the “dark” pathway of dioxetanone intermediate decomposition, it explains the low bioluminescence activity of obelin and apparently of other photoproteins with CTZ-v.

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Section: Discussionmentioning

confidence: 74%

Crystal structure of semi-synthetic obelin-v after calcium induced bioluminescence implies coelenteramine as the main reaction product

Natashin

Eremeeva

Shevtsov

et al. 2022

Sci Rep

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Brief History of Ctenophora

Moroz

2024

Methods in Molecular Biology

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Design of Ctenophore Ca2+-Regulated Photoprotein Berovin Capable of Being Converted into Active Protein Under Physiological Conditions: Computational and Experimental Approaches

Burakova,

Ivanisenko,

Rukosueva

et al. 2024

Life

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Here, we describe (1) the AlphaFold-based structural modeling approach to identify amino acids of the photoprotein berovin that are crucial for coelenterazine binding, and (2) the production and characterization of berovin mutants with substitutions of the identified residues regarding their effects on the ability to form an active photoprotein under physiological conditions and stability to light irradiation. The combination of mutations K90M, N107S, and W103F is demonstrated to cause a shift of optimal conditions for the conversion of apo-berovin into active photoprotein towards near-neutral pH and low ionic strength, and to reduce the sensitivity of active berovin to light. According to the berovin spatial structure model, these residues are found in close proximity to the 6-(p-hydroxy)-phenyl group of the coelenterazine peroxyanion.

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Recombinant light-sensitive photoprotein berovin from ctenophore Beroe abyssicola: Bioluminescence and absorbance characteristics

Cited by 3 publications

References 23 publications

Crystal structure of semi-synthetic obelin-v after calcium induced bioluminescence implies coelenteramine as the main reaction product

Crystal structure of semi-synthetic obelin-v after calcium induced bioluminescence implies coelenteramine as the main reaction product

Brief History of Ctenophora

Design of Ctenophore Ca2+-Regulated Photoprotein Berovin Capable of Being Converted into Active Protein Under Physiological Conditions: Computational and Experimental Approaches

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