Direct evidence for close proximity of catalytic and regulatory domains of heterodimeric sGC based on fluorescence resonance energy transfer
نویسندگان
چکیده
Methods The FRET donor, CFP, and FRET acceptor, YFP, were fused to aminoand carboxy-terminal ends of sGC subunits. After generation of recombinant baculovirus strains fluorescent tagged sGC subunits were co-expressed in Sf9cells. Fluorescent variants of sGC were analyzed in vitro in cytosolic fractions by sensitized emission FRET. In addition, fluorescent tagged sGC subunits were analyzed in vivo using confocal laser scanning microscopy and fluorescence lifetime imaging (FLIM) on an inverted microscope.
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Fluorescent Fusion Proteins of Soluble Guanylyl Cyclase Indicate Proximity of the Heme Nitric Oxide Domain and Catalytic Domain
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