Redox properties of Cys₂His₂ and Cys₄ zinc fingers determined by electrospray ionization mass spectrometry

Abstract

Zinc finger ( ZF ) protein motifs, stabilized by binding of Zn( II ), typically function as interaction modules that bind nucleic acids, proteins and other molecules. The elucidation of the redox states of ZF proteins in cellular conditions, which depend on their midpoint redox potentials, is important for understanding of ZF functioning. In the present study we determined the midpoint redox potentials for representatives of Cys 2 His 2 and Cys 4 types of ZF proteins in apo and Zn( II )‐bound forms using electrospray ionization mass spectrometry. The midpoint redox potentials of the apo forms of Cys 2 His 2 and Cys 4 ZF proteins were −326 and −365 mV ( pH 7.5), respectively. These values are close to the cytosolic redox potential of approx. −350 mV ( pH 7.5) and thus we can conclude that the apo form of Cys 2 His 2 ‐type ZF proteins is predominantly reduced but apo forms of Cys 4 ‐type ZF proteins should be substantially oxidized in the cytoplasm. As expected, Zn( II ) binding stabilized the reduced forms of both ZF proteins: the corresponding redox potential values were −284 and −301 mV , respectively. Consequently, binding of Zn( II ) ions to ZF motifs can act as a sensitive switch that activates the functioning of the ZF motifs within the cell, and also protects them from oxidation and can function as part of a redox‐sensitive regulation mechanism of cellular functions.

Microbial rhodoquinone biosynthesis proceeds via an atypical RquA-catalyzed amino transfer from S-adenosyl-L-methionine to ubiquinone

“…Low activity was also achieved with Zn 2+ in the presence of excess thiol reducing agent. Although zinc is not redox active on its own, it has been shown to facilitate redox chemistry when coordinated with sulfur 30 .…”