aeBlue Chromoprotein Color is Temperature Dependent
Por:
Tamayo-Nunez, Jessica, de la Mora, Javier, Padilla-Vaca, Felipe, Vargas-Maya N.I., Rangel-Serrano, Angeles, Anaya-Velazquez, Fernando, Paramo-Perez, Itzel, Reyes-Martínez J.E., España-Sánchez B.L., Franco, Bernardo
Publicada:
1 ene 2020
Resumen:
BACKGROUND: Marine sessile organisms display a color palette that is the result of the expression of fluorescent and non-fluorescent proteins. Fluorescent proteins have uncovered transcriptional regulation, subcellular localization of proteins, and the fate of cells during development. Chromoproteins have received less attention until recent years as bioreporters. Here, we studied the properties of aeBlue, a a 25.91 kDa protein from the anemone Actinia equina. OBJECTIVE: To assess the properties of aeBlue chromoprotein under different physicochemical conditions. METHODS: In this article, during the purification of aeBlue we uncovered that it suffered a color shift when frozen. We studied the color shift by different temperature incubation and physicochemical conditions and light spectroscopy. To assess the possible structural changes in the protein, circular dichroism analysis, size exclusion chromatography and native PAGE was performed. RESULTS: We uncover that aeBlue chromoprotein, when expressed from a synthetic construct in Escherichia coli, showed a temperature dependent color shift. Protein purified at 4 °C by metal affinity chromatography exhibited a pinkish color and shifts back at higher temperatures to its intense blue color. Circular dichroism analysis revealed that the structure in the pink form of the protein has reduced secondary structure at 4 °C, but at 35 °C and higher, the structure shifts to a native conformation and Far UV- vis CD spectra revealed the shift in an aromatic residue of the chromophore. Also, the chromophore retains its properties in a wide range of conditions (pH, denaturants, reducing and oxidants agents). Quaternary structure is also maintained as a tetrameric conformation as shown by native gel and size exclusion chromatography. CONCLUSION: Our results suggest that the chromophore position in aeBlue is shifted from its native position rendering the pink color and the process to return it to its native blue conformation is temperature dependent. Copyright© Bentham Science Publishers; For any queries, please email at epub@benthamscience.net.
Filiaciones:
Tamayo-Nunez, Jessica:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Univ Guanajuato, Dept Biol, Div Ciencias Nat & Exactas, Noria Alta S-N, Guanajuato 36050, Gto, Mexico
de la Mora, Javier:
Instituto de Fisiologia Celular, Universidad Nacional Autonoma de Mexico, Circuito Exterior S/NMexico City 04510, Mexico
Univ Nacl Autonoma Mexico, Inst Fisiol Celular, Circuito Exterior S-N, Mexico City 04510, DF, Mexico
Padilla-Vaca, Felipe:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Univ Guanajuato, Dept Biol, Div Ciencias Nat & Exactas, Noria Alta S-N, Guanajuato 36050, Gto, Mexico
Vargas-Maya N.I.:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Rangel-Serrano, Angeles:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Univ Guanajuato, Dept Biol, Div Ciencias Nat & Exactas, Noria Alta S-N, Guanajuato 36050, Gto, Mexico
Anaya-Velazquez, Fernando:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Univ Guanajuato, Dept Biol, Div Ciencias Nat & Exactas, Noria Alta S-N, Guanajuato 36050, Gto, Mexico
Paramo-Perez, Itzel:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Univ Guanajuato, Dept Biol, Div Ciencias Nat & Exactas, Noria Alta S-N, Guanajuato 36050, Gto, Mexico
Reyes-Martínez J.E.:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
España-Sánchez B.L.:
CONACYT Centro de Investigacion y Desarrollo Tecnologico en Electroquimica SC. Parque Queretaro s/n Sanfandila, Pedro Escobedo Queretaro. C.P. 76703, Mexico
Franco, Bernardo:
Departamento de Biologia, Division de Ciencias Naturales y Exactas, Universidad de Guanajuato, Noria Alta s/nGuanajuato Gto. 36050, Mexico
Univ Guanajuato, Dept Biol, Div Ciencias Nat & Exactas, Noria Alta S-N, Guanajuato 36050, Gto, Mexico
CONACYT Ctr Invest & Desarrollo Tecnol Electroqui, Parque Queretaro S-N Sanfandila, Pedro Escobedo Queretaro 76703, Mexico
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