Maturation time of GFPmut3 in vitro

Value 7.5 Min Range: Table - link Min
Organism Generic
Reference Iizuka R, Yamagishi-Shirasaki M, Funatsu T. Kinetic study of de novo chromophore maturation of fluorescent proteins. Anal Biochem. 2011 Jul 15 414(2):173-8. p.176 table 1PubMed ID21459075
Method (P.175 left column bottom paragraph:) "The PURE system is a coupled cell-free transcription/ translation system reconstituted from the purified components necessary for translation in E. coli [refs 14,15]." (P.175 right column top paragraph:) "The maturation kinetics was slow enough to monitor in real time with a spectrofluorometer and fitted well to a single exponential curve, suggesting that the oxidation step is the rate-limiting step in the overall chromophore maturation process (Figs. 1 and 2 see also supplementary material). The observed rate constant was defined as the rate constant of de novo chromophore maturation."
Comments "During earlier years, the maturation rate of GFP was determined as the rate of fluorescence development after admission of air to anaerobically expressed GFP [ref 5]. Currently, conventional methods to study the maturation kinetics are based on triggering protein folding of urea-solubilized inclusion bodies [ref 9] and reoxidation of the chromophore after chemical reduction of the mature chromophore [refs 9–13]. However, inconsistent data have been reported (Table 1)."
Entered by Uri M
ID 107004