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Real‐Time Monitoring of the Binding/Dissociation and Redox States of a Single Transition Metal Ions
정승룡,이상욱,홍성철 대한화학회 2018 Bulletin of the Korean Chemical Society Vol.39 No.5
Fluorescence resonance energy transfer (FRET) between organic fluorophores is conventionally used to monitor binding/dissociation and conformational change of macromolecules. Here we use FRET between a cyanine dye and single transition metal ions (tmFRET) to monitor the binding/dissociation of transition metal ions to a synthetic polypeptide, and show that different transition metal ion species can be clearly distinguished based on their binding lifetimes and FRET efficiencies. Furthermore, we demonstrate that the redox state of cytochrome c can be successfully monitored at the single?ion level using the tmFRET.
정승룡,김은지,신수철,송지준,홍성철 대한화학회 2018 Bulletin of the Korean Chemical Society Vol.39 No.2
Prokaryotic Argonaute facilitates the target recognition process by the guide strand via a still unknown mechanism. Using single-molecule fluorescence resonance energy transfer and systematic mutagenesis of Thermus thermophilus Argonaute and guide-target base pairing, we study the kinetic roles of various structural features of guide strand in the prokaryotic Argonaute. We reveal that the 5′-end anchoring of the guide strand, and the sequence complementarity in the seed- and mid-regions greatly accelerate the target binding, and that this acceleration is due to the reduction of the entropic barrier of the binding process. Our data further suggest that the dynamic anchoring of the guide strand to the PAZ domain, which is coupled to target dissociation, does not accompany the partial unwinding of the guide-target duplex.