Abstract
The guanidine-thiocyanate-induced denaturation-renaturation of sfGFP was studied. It was shown that the disruption of sfGFP native structure occurs in the range of guanidine thiocyanate concentrations from 0.5 to 2.5 M. This process was accompanied by simultaneous changes of all recorded parameters. It was found that the small guanidine thiocyanate concentrations (less then 0.1-0.2 M) triggered local structural disturbances in protein which result in significant decrease of chromophore and tryptophan fluorescence intensity and change of protein visible absorption spectrum.
Highlights
A broad variety of fluorescent proteins (FPs) and their engineered analogs are currently used as fluorescent tags of different color’s in bioimaging [1,2,3,4,5]
Encoded FP-containing biosensors based on fluorescence resonance energy transfer (FRET) have been widely adopted for the investigation of protein-protein interactions and other biologically relevant events occurring in the living cell [1, 13]
Visible absorption spectra of sfGFP demonstrated pronounced drop of absorption band corresponding to the anionic form of chromophore with concomitant rise of absorption band corresponding to neutral chromophore
Summary
A broad variety of fluorescent proteins (FPs) and their engineered analogs are currently used as fluorescent tags of different color’s in bioimaging [1,2,3,4,5]. The dependences of different fluorescent characteristics of sfGFP on GTC concentration were recorded after protein incubation in the solution of appropriate concentration at 23◦C during 2, 24, 45, 69, and 95 h.
Sign-in/Register to view highlights & summary Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
