First crystal structures of wild-type and enhanced GFP. Tsien designed T203Y mutant based on crystal structure of S65T GFP. It is yellow fluorescent.
References: M Ormo, AB Cubitt, K Kallio, LA Gross, RY Tsien, SJ Remington: Crystal structure of the Aequorea victoria green fluorescent protein. Science 273 (1996) 1392-95. F Yang, LG Moss, GN Phillips: The molecular structure of green fluorescent protein. Nature Biotechnology 14 (1996) 1246-51. Further reading on Crystal Structures: To Be Added
Posttranslational oxidation of S65T GFP four times faster than wt-GFP. The origin of enhanced GFP (EGFP).
Reference: R Heim, A Cubitt, RY Tsien: Improved green fluorescene. Nature 373 (1995) 663-64.
GFP expressed in E. coli and C. elegans. Mechanism for chromophore formation. First new color (blue) and oxygen dependence reported. Possibility of use in FRET experiments mentioned.
References: M Chalfie, Y Tu, G Euskirchen, WW Ward, DC Prasher: Green fluorescent protein as a marker for gene expression. Science 263 (1994) 802-05. S Inouye, F Tsuji: Evidence for redox forms of the Aequorea green fluorescent protein. FEBS Letters 351 (1994) 211-14. R Heim, DC Prasher, RY Tsien: Wavelength mutations and posttranslational autoxidation of green fluorescent protein. Proc. Natl. Acad. Sci. USA 91 (1994) 12501-04.
Structure of GFP chromophore confirmed, flanking amino acid residues corrected from Shimomura’s 1979 structure. Still “apoGFP”
CW Cody, DC Prasher, WM Westler, FG Pendergast, WW Ward: Chemical Structure of the hexapeptide chromophore of the Aequorea Green fluorescent protein. Biochemistry 32 (1993) 1212-18.
GFP cloned. “These results will enable us to construct an expression vector for the preparation of non-fluorescent apoGFP.”
Reference: DC Prasher, VK Eckenrode, WW Ward, FG Pendergast, MJ Cormier: Primary structure of the Aequorea victorea green fluorescent protein. Gene 111 (1992) 229-33.
Prasher clones and expresses aequorin
Reference: D Prasher, RO McCann, MJ Cormier: Cloning and Expression of the Cdna Coding for Aequorin, a Bioluminescent Calcium-Binding Protein. Biochemical and Biophysical Research Communications 126 (1985) 1259-68.
Intermolecular energy transfer between aequorin and GFP in jellyfish and when co-absorbed on Sephadex column.
“Green protein” named green fluorescent protein.
Reference: JW Hasting, JG Morin: Comparative biochemistry of calcium-activated photoproteins from the ctenophore, Mnemiopsis, and the coelenterates Aequorea, Obelia, Pelagia and Renilla. Biol. Bull. 137 (1969) 402.
GFP identified as protein, extracted from 10,000 jellyfish – “a protein giving solutions that look slightly greenish in sunlight though only yellowish under tungsten lights, and exhibiting a very bright, greenish fluorescence in the ultraviolet of a Mineralite, has also been isolated from squeezates.” Called “green protein.”
Reference: O Shimomura, FH Johnson, Y Saiga: Extraction, purification and properties of aequorin, a bioluminescent protein from the luminous hydromedusan, Aequorea. J. Cell. Comp. Physiol. 59 (1962) 223-29.