GFP Family: Structural Insights into Spectral Tuning

Пахомов, А. А.; Martynov, Vladimir I.

doi:10.1016/j.chembiol.2008.07.009

Cited by 186 publications

(158 citation statements)

References 88 publications

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“…For instance, mCherry is characterized by a particularly distorted chromophore and a rather low fluorescence quantum yield (0.22) [7]. Chromophore deviation from coplanarity can be quantified by the twist around t and f (see Table 2), or also including the tilt angle defined as the deviation of the phenol oxygen from the perfectly planar chromophore [9].…”

Section: Proteins With Rfp Chromophorementioning

confidence: 99%

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One-Photon and Two-Photon Excitation of Fluorescent Proteins

Nifosı̀

Tozzini

2011

Springer Series on Fluorescence

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Fluorescent proteins (FPs) offer a wide palette of colors for imaging applications. One purpose of this chapter is to review the variety of FP spectral properties, with a focus on their structural basis. Fluorescence in FPs originates from the autocatalytically formed chromophore. Several studies exist on synthetic chromophore analogs in gas phase and in solution. Together with the X-ray structures of many FPs, these studies help to understand how excitation and emission energies are tuned by chromophore structure, protonation state, and interactions with the surrounding environment, either solvent molecules or amino acids residues. The increasing use of FPs in two-photon microscopy also prompted detailed investigations of their two-photon excitation properties. The comparison with one-photon excitation reveals nontrivial features, which are relevant both for their implications in understanding multiphoton properties of fluorophores and for application purposes.

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Section: Proteins With Rfp Chromophorementioning

confidence: 99%

“…Several other reviews on FP optical properties are available, some treating more exhaustively the variety of GFP-like fluoro and chromoproteins, and other more focused on application purposes. For recent surveys, see [7][8][9].…”

Section: Introductionmentioning

confidence: 99%

One-Photon and Two-Photon Excitation of Fluorescent Proteins

Nifosı̀

Tozzini

2011

Springer Series on Fluorescence

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“…Fluorescent proteins (FP) Fluorescent proteins are a family of homologous proteins from various organisms, whose major characteristic is to display bright fluorescence in entire visible spectrum (Zhang et al 2002;Pakhomov and Martynov 2008;Okumoto 2010). Green fluorescent protein (GFP), isolated from Aequorea victoria, is the best known fluorescent protein (Tsien 1998).…”

Section: Components Of a Fret Sensormentioning

confidence: 99%

FRET-based genetically-encoded sensors for quantitative monitoring of metabolites

2015

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Neighboring cells in the same tissue can exist in different states of dynamic activities. After genomics, proteomics and metabolomics, fluxomics is now equally important for generating accurate quantitative information on the cellular and sub-cellular dynamics of ions and metabolite, which is critical for functional understanding of organisms. Various spectrometry techniques are used for monitoring ions and metabolites, although their temporal and spatial resolutions are limited. Discovery of the fluorescent proteins and their variants has revolutionized cell biology. Therefore, novel tools and methods targeting sub-cellular compartments need to be deployed in specific cells and targeted to sub-cellular compartments in order to quantify the target-molecule dynamics directly. We require tools that can measure cellular activities and protein dynamics with sub-cellular resolution. Biosensors based on fluorescence resonance energy transfer (FRET) are genetically encoded and hence can specifically target sub-cellular organelles by fusion to proteins or targetted sequences. Since last decade, FRET-based genetically encoded sensors for molecules involved in energy production, reactive oxygen species and secondary messengers have helped to unravel key aspects of cellular physiology. This review, describing the design and principles of sensors, presents a database of sensors for different analytes/processes, and illustrate examples of application in quantitative live cell imaging.

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“…Particularly, some proteins have attracted a more general interest, e.g., green uorescent proteins (GFPs), which are being extensively used as reporters in characterization of protein complexes, 12 expression proles and cell imaging. 13 Due to the intrinsic characteristics of the bacterial envelope, the modied alkaline procedure has become the most frequently explored method for cell lysis in order to extract pDNA.…”

Section: Introductionmentioning

confidence: 99%

Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip

Matos

Senkbeil

Mendonça

et al. 2013

Analyst

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Due to the extensive use of nucleic acid and protein analysis of bacterial samples, there is a need for simple and rapid extraction protocols for both plasmid DNA and RNA molecules as well as reporter proteins like the green fluorescent protein (GFP). In this report, an electropermeability technique has been developed which is based on exposing E. coli cells to low voltages to allow extraction of nucleic acids and proteins.The flow-through electropermeability chip used consists of a microfluidic channel with integrated gold electrodes that promote cell envelope channel formation at low applied voltages. This will allow small biomolecules with diameters less than 30 A to rapidly diffuse from the permeabilized cells to the surrounding solution. By controlling the applied voltage, partial and transient to complete cell opening can be obtained. By using DC voltages below 0.5 V, cell lysis can be avoided and the transiently formed pores can be closed again and the cells survive. This method has been used to extract RNA and GFP molecules under conditions of electropermeability. Plasmid DNA could be recovered when the applied voltage was increased to 2 V, thus causing complete cell lysis.

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GFP Family: Structural Insights into Spectral Tuning

Cited by 186 publications

References 88 publications

One-Photon and Two-Photon Excitation of Fluorescent Proteins

One-Photon and Two-Photon Excitation of Fluorescent Proteins

FRET-based genetically-encoded sensors for quantitative monitoring of metabolites

Nucleic acid and protein extraction from electropermeabilized E. coli cells on a microfluidic chip

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