Fluorescently Labelled ATP Analogues for Direct Monitoring of Ubiquitin Activation
Abstract: Simple and robust assays to monitor enzymatic ATP cleavage with high efficiency in real‐time are scarce. To address this shortcoming, we developed fluorescently labelled adenosine tri‐, tetra‐ and pentaphosphate analogues of ATP. The novel ATP analogues bear — in contrast to earlier reports — only a single acridone‐based dye at the terminal phosphate group. The dye's fluorescence is quenched by the adenine component of the ATP analogue and is restored upon cleavage of the phosphate chain and dissociation of the dye from the adenosine moiety. Thereby the activity of ATP‐cleaving enzymes can be followed in real‐time. We demonstrate this proficiency for ubiquitin activation by the ubiquitin‐activating enzymes UBA1 and UBA6 which represents the first step in an enzymatic cascade leading to the covalent attachment of ubiquitin to substrate proteins, a process that is highly conserved from yeast to humans. We found that the efficiency to serve as cofactor for UBA1/UBA6 very much depends on the length of the phosphate chain of the ATP analogue: triphosphates are used poorly while pentaphosphates are most efficiently processed. Notably, the novel pentaphosphate‐harbouring ATP analogue supersedes the efficiency of recently reported dual‐dye labelled analogues and thus, is a promising candidate for broad applications.
- Location
-
Deutsche Nationalbibliothek Frankfurt am Main
- Extent
-
Online-Ressource
- Language
-
Englisch
- Bibliographic citation
-
Fluorescently Labelled ATP Analogues for Direct Monitoring of Ubiquitin Activation ; volume:26 ; number:28 ; year:2020 ; pages:6279-6284 ; extent:6
Chemistry - a European journal ; 26, Heft 28 (2020), 6279-6284 (gesamt 6)
- Creator
-
Hammler, Daniel
Stuber, Katrin
Offensperger, Fabian
Scheffner, Martin
Zumbusch, Andreas
Marx, Andreas
- DOI
-
10.1002/chem.202001091
- URN
-
urn:nbn:de:101:1-2022060413422922380810
- Rights
-
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Last update
-
15.08.2025, 7:37 AM CEST
Data provider
Deutsche Nationalbibliothek. If you have any questions about the object, please contact the data provider.
Associated
- Hammler, Daniel
- Stuber, Katrin
- Offensperger, Fabian
- Scheffner, Martin
- Zumbusch, Andreas
- Marx, Andreas
Other Objects (12)
Fluorescently Labelled ATP Analogues for Direct Monitoring of Ubiquitin Activation
Fluorescence-Lifetime-Sensitive Probes for Monitoring ATP Cleavage
Aufbau einer Anlage zur Ramanspektroskopie von matrixisolierten Hochtemperaturmolekülen : Untersuchungen an (GeO)n mit n = 1, 2, 3, 4 in festem Argon
Quantifizierung von Apoproteinen - Bedeutung für die Atherosklerose : vergleichende Unters. an e. Normalkollektiv u. Patienten mit e. Hypolipoproteinämie Typ II/Typ IV
Untersuchung zur Eignung einer kombinierten Infusion von Hydroxyethylstärke- und Ringerlösung als Volumenersatz bei der präoperativen Eigenblutspende
Untersuchungen zu den Orientierungseigenschaften des PBLGs und PBDGs
High Sensitivity Stimulated Raman Scattering Microscopy with Electronic Resonance Enhancement
Live Cell Imaging of Enzymatic Turnover of an Adenosine 5′-Tetraphosphate Analog
DNA Conjugation by the Staudinger Ligation : New Thymidine Analogues
Assembly of branched ubiquitin oligomers by click chemistry
Nonlinear optical microscopy with vibrational contrast
Total-internal-reflection fluorescence microscopy with W-shaped axicon mirrors
Fluorescently Labelled ATP Analogues for Direct Monitoring of Ubiquitin Activation
Fluorescence-Lifetime-Sensitive Probes for Monitoring ATP Cleavage
Aufbau einer Anlage zur Ramanspektroskopie von matrixisolierten Hochtemperaturmolekülen : Untersuchungen an (GeO)n mit n = 1, 2, 3, 4 in festem Argon
Quantifizierung von Apoproteinen - Bedeutung für die Atherosklerose : vergleichende Unters. an e. Normalkollektiv u. Patienten mit e. Hypolipoproteinämie Typ II/Typ IV
Untersuchung zur Eignung einer kombinierten Infusion von Hydroxyethylstärke- und Ringerlösung als Volumenersatz bei der präoperativen Eigenblutspende
Untersuchungen zu den Orientierungseigenschaften des PBLGs und PBDGs
High Sensitivity Stimulated Raman Scattering Microscopy with Electronic Resonance Enhancement
Live Cell Imaging of Enzymatic Turnover of an Adenosine 5′-Tetraphosphate Analog
DNA Conjugation by the Staudinger Ligation : New Thymidine Analogues
Assembly of branched ubiquitin oligomers by click chemistry
Nonlinear optical microscopy with vibrational contrast
Total-internal-reflection fluorescence microscopy with W-shaped axicon mirrors
Fluorescently Labelled ATP Analogues for Direct Monitoring of Ubiquitin Activation
Fluorescence-Lifetime-Sensitive Probes for Monitoring ATP Cleavage
Aufbau einer Anlage zur Ramanspektroskopie von matrixisolierten Hochtemperaturmolekülen : Untersuchungen an (GeO)n mit n = 1, 2, 3, 4 in festem Argon
Quantifizierung von Apoproteinen - Bedeutung für die Atherosklerose : vergleichende Unters. an e. Normalkollektiv u. Patienten mit e. Hypolipoproteinämie Typ II/Typ IV
Untersuchung zur Eignung einer kombinierten Infusion von Hydroxyethylstärke- und Ringerlösung als Volumenersatz bei der präoperativen Eigenblutspende
Untersuchungen zu den Orientierungseigenschaften des PBLGs und PBDGs
High Sensitivity Stimulated Raman Scattering Microscopy with Electronic Resonance Enhancement
Live Cell Imaging of Enzymatic Turnover of an Adenosine 5′-Tetraphosphate Analog
DNA Conjugation by the Staudinger Ligation : New Thymidine Analogues
Assembly of branched ubiquitin oligomers by click chemistry
Nonlinear optical microscopy with vibrational contrast