Identification and characterization of fluorescent protein from marine organisms and potentially applications

Abstract

In this study, two GFP from two species of Cnidaria: Aequorea coerulescens and Anemonia sulcata, have been characterized according to the UV – Vis spectroscopy investigations. The goal has been achieved evaluating their fluorescence behaviour in presence of potentially toxic metal ions. A. coerulescens was considered the control system to compare results obtained on the GFP that was extracted from A. sulcata and was partially purified. For the rAcGFP, the small quenching effects in the emission spectrum in the observed conditions suggested that the fluorophore had a low capacity to discriminate the presence of different metals. Different results was obtained for AsGFP about the evolution and the value of quenching effect on the fluorescent emission for all metals used. In fact, the metal – protein bond seemed to be selective. The bond effect changed with the different metals investigated. The order of quenching on the emission protein was Cu(II)˃Pb(II)˃Cd(II).

In questo studio, due GFP provenienti da due distinte specie di Cnidaria: Aequorea coerulescens e Anemonia sulcata, sono state caratterizzate in accordo con la spettroscopia Uv –Vis. L’obiettivo consisteva nel valuare il comportamento della fluorescenza di tali proteine in presenza di metalli potenzialmente tossici. A. coerulescens è stata considerata il sistema controllo a cui comparare i risultati ottenuti sulla GFP estratta e parzialmente purificata da A. sulcata. Per rAcGFP, il limitato effetto di quenching sull’emissione, nelle condizioni osservate suggeriva la scarsa capacità del fluoroforo di discriminare i metalli investigati. Differenti risultati sono stati ottenuti per AsGFP circa l’andamento ed il valore del quenching sull’emissione in fluorescenza, in presenza dei metalli usati. Infatti, il legame metallo – proteina appariva selettivo. L’effetto del legame cambiava con i metalli investigati. L’ordine di quenching sull’emissione della proteina era Cu(II)˃Pb(II)˃Cd(II).