Artículo
Protein crosslinking improves the thermal resistance of plastocyanin immobilized on a modified gold electrode
Autor/es | Olloqui Sariego, José Luis
![]() ![]() ![]() ![]() ![]() ![]() ![]() Díaz Quintana, Antonio Jesús ![]() ![]() ![]() ![]() ![]() ![]() ![]() Rosa Acosta, Miguel Ángel de la ![]() ![]() ![]() ![]() ![]() ![]() Calvente Pacheco, Juan José ![]() ![]() ![]() ![]() ![]() ![]() ![]() Márquez Escudero, Inmaculada Díaz Moreno, Irene ![]() ![]() ![]() ![]() ![]() ![]() ![]() Andreu Fondacabe, Rafael Jesús ![]() ![]() ![]() ![]() ![]() ![]() ![]() |
Departamento | Universidad de Sevilla. Departamento de Química Física Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular |
Fecha de publicación | 2018-12 |
Fecha de depósito | 2024-01-30 |
Publicado en |
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Resumen | Increasing the thermal stability of immobilized proteins is a motivating goal for improving the performance of electrochemical biodevices. In this work, we propose the immobilization of crosslinked plastocyanin from the ... Increasing the thermal stability of immobilized proteins is a motivating goal for improving the performance of electrochemical biodevices. In this work, we propose the immobilization of crosslinked plastocyanin from the thermophilic cyanobacterium Phormidium laminosum by simultaneous incubation of a mixture of plastocyanin and the coupling reagents. The thermal stability of the so built covalently immobilized protein films has been assessed by cyclic voltammetry in the 0–90 °C temperature range and has been compared to that of physisorbed films. It is shown that the protein loss along a thermal cycle is significantly reduced in the case of the crosslinked films, whose redox properties remain unaltered along a cyclic heating-cooling thermal scan, and can withstand the contact with 70 °C solutions for four hours. Comparison of thermal unfolding curves obtained by circular dichroism spectroscopy of both free and crosslinked protein confirms the improved thermic resistance of the crosslinked plastocyanin. Notably, the electron transfer thermodynamics of physisorbed and crosslinked plastocyanin films are quite similar, suggesting that the formation of intra- and inter-protein amide bonds do not affect the integrity and functionality of the copper redox centers. UV–Vis absorption and circular dichroism measurements corroborate that protein crosslinking does not alter the coordination geometry of the metal center. |
Agencias financiadoras | Ministerio de Economía y Competitividad (MINECO). España European Commission (EC). Fondo Europeo de Desarrollo Regional (FEDER) |
Identificador del proyecto | CTQ2014-52641-P
![]() CTQ2015-71955-REDT ![]() BFU2015-71017/BMC ![]() |
Cita | Olloqui Sariego, J.L., Díaz Quintana, A.J., Rosa Acosta, M.Á.d.l., Calvente Pacheco, J.J., Márquez Escudero, I., Díaz Moreno, I. y Andreu Fondacabe, R.J. (2018). Protein crosslinking improves the thermal resistance of plastocyanin immobilized on a modified gold electrode. Bioelectrochemistry, 124, 127-132. https://doi.org/10.1016/j.bioelechem.2018.07.013. |
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10.1016_j.bioelechem.2018.07.0 ... | 1.341Mb | ![]() | Ver/ | Versión Aceptada |