Autor(es):Mostrar menos -

Soares, Douglas M. M. ; Galeazzo, Gabriela A. ; Sgro, German G. ; de Moraes, Gabriela V. ; Kronenberg, Leora ; Borukh, Emmanuella ; Migotto, Alvaro E. ; Gruber, David F. ; Sparks, John S. ; Pieribone, Vincent A. ; Stevani, Cassius V. ; Oliveira, Anderson G.

• Tipo de documento: Artigo Científico
• Fonte: FEBS Journal; v. N/A, p. 17-pg., 2025-04-18.

Resumo

Ca2+-regulated photoproteins (CaPhs) consist of single-chain globular proteins to which coelenterazine, a widely distributed marine luminogenic substrate (the luciferin), binds along with molecular oxygen, producing a stable peroxide. Upon Ca2+ addition, CaPhs undergo conformational changes leading to the cyclization of the peroxide and the formation of a high-energy intermediate. Subsequently, its decomposition yields coelenteramide in an excited state and results in the emission of a flash of light. To date, most known CaPh systems emit blue light (lambda max 465-495 nm), except for two bolinopsin isospecies that emit green light (lambda max 500 nm). Here, we report the cloning and functional characterization of wild-type CaPhs capable of emitting green light: velamins, isolated from the bioluminescent ctenophore Velamen parallelum. Ten unique photoprotein-like sequences were recovered and grouped in three main clusters. Representative sequences were cloned, expressed, purified, and regenerated into the active His-tagged alpha-, beta-, and gamma-velamins. Upon injection of a calcium-containing buffer into the velamin, a flash of green light (lambda max 500-508 nm) was observed across pH values ranging from 7 to 9. Whilst alpha-velamin isoforms exhibited the highest light emission activity, beta- and gamma-velamins were found to be more thermostable at higher temperatures. Velamins are the wild-type CaPhs with the longest-wavelength light emission yet reported, making them an excellent model for investigating spectral modulation mechanisms in photoproteins. (AU)

• Processo FAPESP: 19/25086-1 - Estudo do processo de bioluminescência em Harmothoe sp., Kinberg, 1856 e suas funções ecológicas
• Beneficiário: Gabriela Verruck de Moraes
• Modalidade de apoio: Bolsas no Brasil - Doutorado Direto
• Processo FAPESP: 17/22501-2 - Quimiexcitação eletrônica em sistemas biológicos: bioluminescência e 'foto'química no escuro
• Beneficiário: Etelvino José Henriques Bechara
• Modalidade de apoio: Auxílio à Pesquisa - Temático
• Processo FAPESP: 19/12605-0 - Bioluminescência de Fungos, Centopeia e Organismos Marinhos: Aspectos Químicos e Biológicos
• Beneficiário: Douglas Moraes Mendel Soares
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 20/11189-0 - Estudos estruturais e funcionais dos sistemas de secreção de bactérias usando crio-microscopia eletrônica
• Beneficiário: Germán Gustavo Sgro
• Modalidade de apoio: Auxílio à Pesquisa - Regular
• Processo FAPESP: 22/14964-0 - Taxonomia molecular de fungos bioluminescentes brasileiros e bioluminescência do milípede Motyxia sequoia
• Beneficiário: Douglas Moraes Mendel Soares
• Modalidade de apoio: Bolsas no Exterior - Estágio de Pesquisa - Pós-Doutorado