RoGFP (original) (raw)
roGFP (englisch Redox-sensitive GFP) ist eine veränderte Variante des grün fluoreszierenden Proteins, bei der sich die Fluoreszenz redox-abhängig ändert.
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| dbo:abstract | roGFP (englisch Redox-sensitive GFP) ist eine veränderte Variante des grün fluoreszierenden Proteins, bei der sich die Fluoreszenz redox-abhängig ändert. (de) The reduction-oxidation sensitive green fluorescent protein (roGFP) is a green fluorescent protein engineered to be sensitive to changes in the local redox environment. roGFPs are used as redox-sensitive biosensors. In 2004, researchers in 's lab at the University of Oregon constructed the first roGFPs by introducing two cysteines into the beta barrel structure of GFP. The resulting engineered protein could exist in two different oxidation states (reduced dithiol or oxidized disulfide), each with different fluorescent properties. Originally, members of the Remington lab published six versions of roGFP, termed roGFP1-6 (see more structural details below). Different groups of researchers introduced cysteines at different locations in the GFP molecule, generally finding that cysteines introduced at the amino acid positions 147 and 204 produced the most robust results. roGFPs are often genetically encoded into cells for in-vivo imaging of redox potential. In cells, roGFPs can generally be modified by redox enzymes such as glutaredoxin or thioredoxin. roGFP2 preferentially interacts with glutaredoxins and therefore reports the cellular glutathione redox potential. Various attempts have been made to make roGFPs that are more amenable to live-cell imaging. Most notably, substituting three positively-charged amino acids adjacent to the disulfide in roGFP1 drastically improves the response rate of roGFPs to physiologically relevant changes in redox potential. The resulting roGFP variants, named roGFP1-R1 through roGFP1-R14, are much more suitable for live-cell imaging. The roGFP1-R12 variant has been used to monitor redox potential in bacteria and yeast, but also for studies of spatially-organized redox potential in live, multicellular organisms such as the model nematode C. elegans. In addition, roGFPs are used to investigate the topology of ER proteins, or to analyze the ROS production capacity of chemicals. One notable improvement to roGFPs occurred in 2008, when the specificity of roGFP2 for glutathione was further increased by linking it to the human glutaredoxin 1 (Grx1). By expressing the Grx1-roGFP fusion sensors in the organism of interest and/or targeting the protein to a cellular compartment, it is possible to measure the glutathione redox potential in a specific cellular compartment in real-time and therefore provides major advantages compared to other invasive static methods e.g. HPLC. Given the variety of roGFPs, some effort has been made to benchmark their performance. For example, members of 's group published a method in 2020 describing the 'suitable ranges' of different roGFPs, determined by how sensitive each sensor is to experimental noise in different redox conditions. (en) |
| dbo:pdb | 1JC1 |
| dbo:symbol | roGFP |
| dbo:thumbnail | wiki-commons:Special:FilePath/RoGFP1-R7_Oxidized_and_Reduced.png?width=300 |
| dbo:wikiPageID | 22064427 (xsd:integer) |
| dbo:wikiPageLength | 11999 (xsd:nonNegativeInteger) |
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| dbo:wikiPageWikiLink | dbr:Endoplasmic_reticulum dbr:Synapto-pHluorin dbc:Fluorescent_proteins dbr:Glutathione dbr:Redox_potential dbr:Thioredoxin dbr:C._elegans dbr:Dithiol dbr:Amino_acid dbc:Cell_imaging dbr:Cysteine dbr:Fluorescent dbc:Biochemistry_detection_methods dbr:Glutaredoxin dbr:Protein dbr:Redox dbr:High-performance_liquid_chromatography dbr:Biosensor dbc:Biosensors dbr:Disulfide dbr:Green_fluorescent_protein dbr:Glutaredoxins dbr:Reactive_oxygen_species dbr:Chemicals dbr:Javier_Apfeld dbr:S._James_Remington |
| dbp:caption | The oxidized and reduced forms of the redox-sensitive Green Fluorescent Protein 1-R7 . (en) |
| dbp:name | reduction-oxidation sensitive Green fluorescent protein (en) |
| dbp:pdb | 1 (xsd:integer) |
| dbp:symbol | roGFP (en) |
| dbp:width | 400 (xsd:integer) |
| dbp:wikiPageUsesTemplate | dbt:Reflist dbt:Short_description dbt:Infobox_protein |
| dcterms:subject | dbc:Fluorescent_proteins dbc:Cell_imaging dbc:Biochemistry_detection_methods dbc:Biosensors |
| gold:hypernym | dbr:Biosensor |
| rdf:type | owl:Thing dbo:Biomolecule wikidata:Q206229 wikidata:Q8054 yago:WikicatBiochemistryMethods yago:Ability105616246 yago:AbsorptionIndicator114598383 yago:Abstraction100002137 yago:Cognition100023271 yago:ColoringMaterial114984973 yago:Dye114985383 yago:Fluorescein114987695 yago:Indicator114917208 yago:Know-how105616786 yago:Material114580897 yago:Matter100020827 yago:Method105660268 yago:Part113809207 yago:PhysicalEntity100001930 yago:PsychologicalFeature100023100 yago:Relation100031921 dbo:Protein yago:Substance100019613 yago:WikicatFluorescentDyes |
| rdfs:comment | roGFP (englisch Redox-sensitive GFP) ist eine veränderte Variante des grün fluoreszierenden Proteins, bei der sich die Fluoreszenz redox-abhängig ändert. (de) The reduction-oxidation sensitive green fluorescent protein (roGFP) is a green fluorescent protein engineered to be sensitive to changes in the local redox environment. roGFPs are used as redox-sensitive biosensors. In 2004, researchers in 's lab at the University of Oregon constructed the first roGFPs by introducing two cysteines into the beta barrel structure of GFP. The resulting engineered protein could exist in two different oxidation states (reduced dithiol or oxidized disulfide), each with different fluorescent properties. (en) |
| rdfs:label | RoGFP (de) RoGFP (en) |
| owl:sameAs | freebase:RoGFP yago-res:RoGFP wikidata:RoGFP dbpedia-de:RoGFP https://global.dbpedia.org/id/4uFat |
| prov:wasDerivedFrom | wikipedia-en:RoGFP?oldid=1093287912&ns=0 |
| foaf:depiction | wiki-commons:Special:FilePath/RoGFP1-R7_Oxidized_and_Reduced.png |
| foaf:isPrimaryTopicOf | wikipedia-en:RoGFP |
| is dbo:wikiPageRedirects of | dbr:Redox-sensitive_Green_Fluorescent_Protein dbr:Redox-sensitive_green_fluorescent_protein dbr:Redox_imaging |
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| is foaf:primaryTopic of | wikipedia-en:RoGFP |
