Structure–Function Relationships in Fluorescent Marker Proteins of the Green Fluorescent Protein Family

2011 ◽  
pp. 241-263 ◽  
Author(s):  
G. Ulrich Nienhaus ◽  
Karin Nienhaus ◽  
Jörg Wiedenmann
Keyword(s):  
Green Fluorescent Protein ◽  
Structure Function ◽  
Fluorescent Protein ◽  
Protein Family ◽  
Fluorescent Marker ◽  
Marker Proteins ◽  
Green Fluorescent

scholarly journals Use of a Transient Assay for Studying the Genetic Determinants of Fv1 Restriction

2000 ◽  
Vol 74 (16) ◽  
pp. 7422-7430 ◽  
Author(s):  
Michael Bock ◽  
Kate N. Bishop ◽  
Greg Towers ◽  
Jonathan P. Stoye
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Leukemia Virus ◽  
Yellow Fluorescent Protein ◽  
Retroviral Vectors ◽  
Genetic Determinants ◽  
Entry Site ◽  
Protein Coding ◽  
Fluorescent Marker ◽  
Green Fluorescent

ABSTRACT To probe the genetic determinants controlling the interaction between the retroviral restriction gene Fv1 and its murine leukemia virus target, we set out to develop rapid, transient assays for Fv1 function. Cells were transfected or transduced withFv1 expression plasmids which can produce green fluorescent protein via an internal ribosome entry site positioned between theFv1 and green fluorescent protein coding sequences.Fv1 function was then assessed by comparing virus replication in green fluorescent protein-positive and -negative cells, using retroviral vectors encoding a second fluorescent marker, yellow fluorescent protein, or β-galactosidase. Using this assay, we could show that Fv1 specificities were not as absolute as previously thought, since the Fv1b allele was capable of interacting with “nonrestricted” B- and NB-tropic viruses and by shuffling the n- and b-alleles of Fv1, it was possible to generate a Fv1 molecule capable of restricting N-, B-, and NB-tropic viruses equally efficiently. Further, we could show that the presence of nonrestricting Fv1 in the same cell as restrictive Fv1 abrogates restriction, implying competition for binding to the retroviral target.

Fluorophore Conformation in Green Fluorescent Protein: A Quantum Mechanics/Molecular Mechanics Study

2004 ◽  
Vol 826 ◽  
Author(s):  
Steven Trohalaki ◽  
Soumya S. Patnaik ◽  
Ruth Pachter
Keyword(s):  
Quantum Mechanics ◽  
Green Fluorescent Protein ◽  
Molecular Mechanics ◽  
Fluorescent Protein ◽  
Protein A ◽  
Fluorescent Marker ◽  
Torsional Potential ◽  
Central Helix ◽  
Protein Cavity ◽  
Green Fluorescent

AbstractGreen Fluorescent Protein (GFP) is a widely used fluorescent marker exhibiting two excitation peaks – a strong peak at 398 nm and a second at 475 nm, with the fluorescence at ca. 510 nm. Its molecular structure consists of a β-barrel composed of 11 β-strands and a central helix containing the fluorophore. Two different forms of the fluorophore – a protonated/neutral fluorophore and a de-protonated/anionic fluorophore – are thought to be responsible for the two distinct spectroscopic states. Notably, the isolated fluorophore in solution is efficiently quenched. Conformational flexibility within the protein cavity is an implicitly important factor that governs the photochemistry of GFP. However, the literature contains accounts of studies that reach conflicting conclusions, claiming that either the fluorophore's barrier to internal rotation is negligibly small or that the protein cavity is not complementary to a planar fluorophore. In this work, we calculate the torsional potential of one of the two exocyclic bonds that connect the two rings in the fluorophore, taking into account its immediate environment by applying a quantum mechanics/molecular mechanics method, with the ultimate aim of evaluating the protein-environment effects on the fluorescence.

scholarly journals Diversity and evolution of the green fluorescent protein family

2002 ◽  
Vol 99 (7) ◽  
pp. 4256-4261 ◽  
Author(s):  
Y. A. Labas ◽  
N. G. Gurskaya ◽  
Y. G. Yanushevich ◽  
A. F. Fradkov ◽  
K. A. Lukyanov ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Protein Family ◽  
Green Fluorescent

scholarly journals The key to the yellow-to-cyan tuning in the green fluorescent protein family is polarisation

2019 ◽  
Vol 21 (35) ◽  
pp. 18988-18998 ◽  
Author(s):  
Riccardo Nifosì ◽  
Benedetta Mennucci ◽  
Claudia Filippi
Keyword(s):  
Green Fluorescent Protein ◽  
Excitation Energy ◽  
Fluorescent Protein ◽  
Transition Density ◽  
Protein Family ◽  
Specific Induction ◽  
Green Fluorescent

Within a QM/MM approach, correctly predicting the tuning of the excitation energy of the chromophore in the GFP family requires to account for state-specific induction and for the coupling of the MM polarisation to the chromophore transition density.

scholarly journals Analytical ultracentrifugation as a tool in the studies of aggregation of the fluorescent marker, Enhanced Green Fluorescent Protein

2020 ◽  
Author(s):  
Aleksandra Dawidziak-Pakula ◽  
Joanna Krasowska ◽  
Beata Wielgus-Kutrowska
Keyword(s):  
Green Fluorescent Protein ◽  
Enhanced Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Analytical Ultracentrifugation ◽  
De Novo ◽  
Continuous Distribution ◽  
Fluorescent Marker ◽  
Low Concentrations ◽  
Green Fluorescent ◽  
De Novo Folding

Enhanced green fluorescent protein (EGFP) is a fluorescent marker used in bio-imaging applications, including as an indicator of folding or aggregation of a fused partner. However, the limited maturation, low folding efficiency, and presence of non-fluorescent states of EGFP can influence the interpretation of experimental data. To measure aggregation associated with de novo folding of EGFP from a high GdnHCl concentration, the analytical ultracentrifugation method was used. Absorption detection at 280 nm allowed to monitor the presence of monomers and aggregated forms. Fluorescence detection enabled the observation of only properly folded molecules with a functional chromophore. The results showed intensive aggregation of EGFP in low concentrations of GdnHCl with a continuous distribution of aggregated forms. The properly folded monomers with mature chromophore were fluorescent, while the conglomerates of EGFP molecules were not. These facts are essential for a proper interpretation of data obtained with EGFP labelling.

Spectral diversity among members of the green fluorescent protein family in hydroid jellyfish (Cnidaria, Hydrozoa)

2005 ◽  
Vol 31 (1) ◽  
pp. 43-47 ◽  
Author(s):  
Yu. G. Yanushevich ◽  
D. A. Shagin ◽  
A. F. Fradkov ◽  
K. S. Shakhbazov ◽  
E. V. Barsova ◽  
...  
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Protein Family ◽  
Green Fluorescent

Chromoproteins of the green fluorescent protein family: Properties and applications

2008 ◽  
Vol 34 (5) ◽  
pp. 517-525 ◽  
Author(s):  
M. A. Shkrob ◽  
A. S. Mishin ◽  
D. M. Chudakov ◽  
Yu. A. Labas ◽  
K. A. Lukyanov
Keyword(s):  
Green Fluorescent Protein ◽  
Fluorescent Protein ◽  
Protein Family ◽  
Green Fluorescent
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