| Range |
~60 Table - link Min
|
| Organism |
Bacteria Escherichia coli |
| 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
|
| Primary Source |
J.A. Sniegowski, J.W. Lappe, H.N. Patel, H.A. Huffman, R.M. Wachter, Base catalysis of chromophore formation in Arg96 and Glu222 variants of green fluorescent protein, J. Biol. Chem. 280 (2005) 26248–26255.PubMed ID15888441
|
| Method |
(Footnote 'e' beneath table:) "Rate of GFP prepared from inclusion bodies at 30°C. Values were taken from [primary source]." (Primary source p.26248 right column 2nd paragraph:)"The kinetics of de novo chromophore formation has been
determined by monitoring the rate of fluorescence acquisition.
The GFP-S65T maturation process, inclusive of protein folding
and the ensuing chemical steps, has been shown to proceed in
vitro with a time constant of 122 min [ref 15]. In this experiment,
maturation was induced by rapid dilution of urea-solubilized
inclusion bodies at room temperature. De novo protein folding
was monitored independently by a trypsin resistance assay and
was estimated to proceed with a time constant of 14 min
(t1/2~10 min)." |
| 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 |
107001 |