Optimising the synthesis, polymer membrane encapsulation and photoreduction performance of Ru(II)- and Ir(III)-bis(terpyridine) cytochrome c bioconjugates

\u3cp\u3eRuthenium(ii) and iridium(iii) bis(terpyridine) complexes were prepared with maleimide functionalities in order to site-specifically modify yeast iso-1 cytochrome c possessing a single cysteine residue available for modification (CYS102). Single X-ray crystal structures were solved for aniline and maleimide Ru(ii) 3 and Ru(ii) 4, respectively, providing detailed structural detail of the complexes. Light-activated bioconjugates prepared from Ru(ii) 4 in the presence of tris(2-carboxyethyl)-phosphine (TCEP) significantly improved yields from 6% to 27%. Photoinduced electron transfer studies of Ru(ii)-cyt c in bulk solution and polymer membrane encapsulated specimens were performed using EDTA as a sacrificial electron donor. It was found that membrane encapsulation of Ru(ii)-cyt c in PS\u3csub\u3e140\u3c/sub\u3e-b-PAA\u3csub\u3e48\u3c/sub\u3e resulted in a quantum efficiency of 1.1 ± 0.3 × 10\u3csup\u3e-3\u3c/sup\u3e, which was a two-fold increase relative to the bulk. Moreover, Ir(iii)-cyt c bioconjugates showed a quantum efficiency of 3.8 ± 1.9 × 10\u3csup\u3e-1\u3c/sup\u3e, equivalent to a ∼640-fold increase relative to bulk Ru(ii)-cyt c.\u3c/p\u3