Circularly permuted variants of the green fluorescent protein

@article{Topell1999CircularlyPV,
  title={Circularly permuted variants of the green fluorescent protein},
  author={Simon Topell and Jens Hennecke and Rudi Glockshuber},
  journal={FEBS Letters},
  year={1999},
  volume={457}
}
Circularly permuted monomeric red fluorescent proteins with new termini in the β‐sheet
TLDR
The extensive directed evolution of the variant with new termini at position 193 of the protein sequence for improved fluorescent brightness is reported, known as cp193g7, has 61% of the intrinsic brightness of mCherry and was found to be highly tolerant of circular permutation at other locations within the sequence.
Efficiently folding and circularly permuted variants of the Sapphire mutant of GFP
TLDR
By directed rational mutagenesis, a new variant of the Sapphire mutant of GFP with improved folding properties that turns out to be especially beneficial when expressed within organelles or as a fusion tag is produced.
Cyclic Green Fluorescent Protein Produced in Vivo Using an Artificially Split PI-PfuI Intein from Pyrococcus furiosus *
TLDR
Green fluorescent protein (GFP) was cyclized with this method in vivo on milligram scales and might become a novel tool for studying the role of termini and backbone topology in various biological processes such as protein degradation and translocation in vivo as well as in vitro.
Engineering and characterization of a superfolder green fluorescent protein
TLDR
A robustly folded version of GFP is generated, called 'superfolder' GFP, that folds well even when fused to poorly folded polypeptides, and shows improved tolerance of circular permutation, greater resistance to chemical denaturants and improved folding kinetics.
Specific modification at the C-terminal lysine residue of the green fluorescent protein variant, GFPuv, expressed in Escherichia coli
TLDR
The recombinant expression of G FPuv, a commonly-used mutant of GFP, in E. coli produced two distinct molecular species as judged by in-gel fluorescence SDS-PAGE, which could be separately purified by anion-exchange chromatography without any remarkable differences in the fluorescence spectra.
Functional Characterization of Permuted Enhanced Green Fluorescent Proteins Comprising Varying Linker Peptides ¶
TLDR
Laser‐scanning microscopy of HeLa cells demonstrated that the PEGFP are particularly well suited as fluorescent indicators in two‐photon imaging, and expression dynamics of P EGFP were revealed to be similar to that of EGFP.
Tolerance of a Knotted Near-Infrared Fluorescent Protein to Random Circular Permutation.
TLDR
Results show how the contact order of a knotted BphP can be altered without disrupting chromophore binding and fluorescence, an important step toward the creation of near-infrared biosensors with expanded chemical sensing functions for in vivo imaging.
Functional Characterization of Permuted Enhanced Green Fluorescent Proteins Comprising Varying Linker Peptides¶
TLDR
Laser-scanning microscopy of HeLa cells demonstrated that the PEGFP are particularly well suited as fluorescent indicators in two-photon imaging and expression dynamics of PEG FP were revealed to be similar to that of EGFP.
Identification of Sites Within a Monomeric Red Fluorescent Protein that Tolerate Peptide Insertion and Testing of Corresponding Circular Permutations
TLDR
This work has investigated the tolerance of an engineered monomeric descendent of Discosoma RFP, known as mCherry, towards peptide insertion and circular permutation and identified six genetically distinct sites localized in three different loops where a sequence of five residues could be inserted without abolishing the ability of the protein to form its intrinsic red fluorescent chromophore.
Understanding the folding of GFP using biophysical techniques
TLDR
The recent literature on protein engineering studies that have improved the folding properties of GFP are reviewed and the biophysical work on the folding of G FP is discussed, beginning to reveal how this large and complex structure forms.
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