Fluorescent proteins isolation

Prendergast, F. G., and Mann, K. G. (1978). Chemical and physical properties of aequorin and the green fluorescent protein isolated from Aequorea forskalea. Biochemistry 17 3448-3453. 

An intensely fluorescent protein isolated from the jellyfish Aequorea victoria and other Aequorea species. This protein produces its fluorescence from an intrinsic chro-mophore generated by a series of steps involving residues 65-67 of the polypeptide chain. As a result, GFP has broad application, especially when expressed as a fused or chimeric protein containing a second polypeptide region whose intracellular location is of interest. ... 

Two types of fluorescent proteins have been isolated from luminous bacteria and studied in detail. The first of them are the blue fluorescent lumazine proteins (LumPs) containing lumazine as their chromophores, which were isolated from P. phosphoreum and P. fischeri (Gast and Lee, 1978 Koda and Lee, 1979 O Kane et al.y 1985). The second are the yellow fluorescent proteins (YFPs) containing a chromophore of FMN or riboflavin, isolated from P. fischeri strain Y-l (Daubner et al., 1987 Macheroux et ai, 1987 ... 

Campbell and Herring (1987) isolated and partially purified a red fluorescent protein from the suborbital light organs of M. niger. The absorption spectrum of this red fluorescent protein had a peak at 612 nm, a shoulder at 555 nm, and a secondary peak at 490 nm. 

Its fluorescence showed two emission peaks, 564 nm and 626 nm the excitation peaks for the former emission peak were at 305 nm and 374 nm, and those for the latter were at 332 nm and 392 nm, indicating the presence of two different components in the preparation. It appears that the characteristics of the fluorescent proteins were altered by isolation and purification. 

Levine, L. D., and Ward, W. W. (1982). Isolation and characterization of a photoprotein, phialidin, and a spectrally unique green-fluorescent protein from the bioluminescent jellyfish Phialidium gregarium. Comp. Biochem. Physiol. 72B 77-85. 

Voityuk AA, Michel-Beyerle ME, Rosch N (1998) Structure and rotation barriers for ground and excited states of the isolated chromophore of the green fluorescent protein. Chem Phys Lett 296 269-276... 

The second major breakthrough for the application of fluorescent proteins was the isolation of the red fluorescent protein (RFP) drFP583 or DsRed from the Anthozoa and Discosoma sp., a mushroom-shaped anemone found in the warm waters of the Indo-Pacific ocean [13], The breakthrough was not only the discovery of the first true RFP, but equally important was the fact that it was discovered in a nonbioluminescent species and that the gene was cloned immediately. 

The isolation of fluorescent proteins from nonbioluminescent species has led to the discovery of a super family of GFP-like proteins [1, 14]. Recently, six additional GFP-like proteins were isolated from A. victoria-related jellyfish [14-16]. Furthermore, a large number of GFP-like proteins have been isolated from Anthozoa species, ranging in fluorescence from green to orange-red, as well as nonflu-orescent purple-blue chromoproteins [17-19],... 

Hydrozoa, such as A. victoria, and Anthozoa both belong to the phylum Cnidaria. In addition, fluorescent proteins have been isolated from planktonic Copepods, which belong to the evolutionary distant phylum Arthropodia [14], This wide phylogenetic distribution of GFP-like proteins might implicate that these proteins developed early in evolution and hence that almost every animal taxon can potentially contain GFP homologs [1],... 

TurboFP was also used as a basis for far-red fluorescent proteins (fRFPs). Residues surrounding the chromophore were mutagenized to create a library, which was subsequently subjected to random mutagenesis. A bright far-red variant with excitation and emission maxima at 588 and 635 nm, respectively was isolated and named Katushka [79]. This fast-maturing protein has an... 

Several other fluorescent proteins have been isolated from a variety of species. We will briefly discuss the variants with the most optimal spectroscopic properties. A true orange fluorescent protein was isolated from the stony coral Fungia concinna, with good absorbance and a quantum yield of 0.45. Targeted mutagenesis was used to monomerize the protein, yielding mKO with an absorbance of 51,600 M-1 cm 1 and a quantum yield of 0.74 [84, 85], The crystal... 

A bright cyan-green fluorescent protein was isolated from Clavu-laria coral [86]. Since one of the intermediates displayed fast bleaching, a screen for more photostable variants was performed. The optimized monomeric variant was named teal fluorescent protein 1 (mTFPl). It has an excitation and emission maximum at 462 and 492 nm, respectively, so this protein is spectrally located in between CFP and GFP. With an extinction coefficient of 64,000 M 1 cm-1 and a quantum yield of 0.85 mTFPl is a very bright fluorescent protein. 

Pongor S, Ulrich PC, Bencsath FA and Cerami A (1984) Aging of proteins isolation and identification of a fluorescent chromophore from the reaction of polypeptides with glucose. Proc Natl Acad Sci USA 81, 2684-2688. 

The green fluorescent protein is a specialized protein that has found many applications. Isolated from jellyfish, it has also been obtained by inserting the jellyfish gene in the genome of the tobacco plant (Boevink 1999). However, the process is still far from the efficiency needed by industry. 

Excited-state dynamics in the Green Fluorescent Proteins the cases of wild type, uv-mutant, and isolated synthetic analogues of the GFP chromophore. 

---