Photoinduced electron transfer (PET)-probes for the study of enzyme activity at the ensemble and single molecule level

2007 ◽  
Author(s):  
Sigrun Henkenjohann ◽  
Sebastian van de Linde ◽  
Sören Doose ◽  
Philip Tinnefeld ◽  
Markus Sauer
Keyword(s):  
Electron Transfer ◽  
Enzyme Activity ◽  
Single Molecule ◽  
Photoinduced Electron Transfer ◽  
Single Molecule Level ◽  
Pet Probes

scholarly journals Two-colour single-molecule photoinduced electron transfer fluorescence imaging microscopy of chaperone dynamics

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Jonathan Schubert ◽  
Andrea Schulze ◽  
Chrisostomos Prodromou ◽  
Hannes Neuweiler
Keyword(s):  
Electron Transfer ◽  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Conformational Changes ◽  
Photoinduced Electron Transfer ◽  
Structural Changes ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Fluorescence Imaging Microscopy ◽  
Pet Probes

AbstractMany proteins are molecular machines, whose function is dependent on multiple conformational changes that are initiated and tightly controlled through biochemical stimuli. Their mechanistic understanding calls for spectroscopy that can probe simultaneously such structural coordinates. Here we present two-colour fluorescence microscopy in combination with photoinduced electron transfer (PET) probes as a method that simultaneously detects two structural coordinates in single protein molecules, one colour per coordinate. This contrasts with the commonly applied resonance energy transfer (FRET) technique that requires two colours per coordinate. We demonstrate the technique by directly and simultaneously observing three critical structural changes within the Hsp90 molecular chaperone machinery. Our results reveal synchronicity of conformational motions at remote sites during ATPase-driven closure of the Hsp90 molecular clamp, providing evidence for a cooperativity mechanism in the chaperone’s catalytic cycle. Single-molecule PET fluorescence microscopy opens up avenues in the multi-dimensional exploration of protein dynamics and allosteric mechanisms.

scholarly journals Photoinduced Electron Transfer of Oxazine 1/TiO2Nanoparticles at Single Molecule Level by Using Confocal Fluorescence Microscopy

Langmuir ◽  
2010 ◽  
Vol 26 (11) ◽  
pp. 9050-9060 ◽  
Author(s):  
Yi-Ju Chen ◽  
Hsin-Yu Tzeng ◽  
Hsiu-Fang Fan ◽  
Ming-Shiang Chen ◽  
Jer-Shing Huang ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Photoinduced Electron Transfer ◽  
Confocal Fluorescence Microscopy ◽  
Single Molecule Level ◽  
Confocal Fluorescence ◽  
Oxazine 1

Intramolecular Distances and Dynamics from the Combined Photon Statistics of Single-Molecule FRET and Photoinduced Electron Transfer

2013 ◽  
Vol 117 (42) ◽  
pp. 13015-13028 ◽  
Author(s):  
Dominik Haenni ◽  
Franziska Zosel ◽  
Luc Reymond ◽  
Daniel Nettels ◽  
Benjamin Schuler
Keyword(s):  
Electron Transfer ◽  
Single Molecule ◽  
Photoinduced Electron Transfer ◽  
Photon Statistics ◽  
Single Molecule Fret

Electron Transfer in a Naphthalene Diimide System Studied by Single-Molecule Delayed Fluorescence

2020 ◽  
Vol 73 (8) ◽  
pp. 699
Author(s):  
Rosalind P. Cox ◽  
Saman Sandanayake ◽  
Steven J. Langford ◽  
Toby D. M. Bell
Keyword(s):  
Electron Transfer ◽  
Single Molecule ◽  
Delayed Fluorescence ◽  
Pmma Film ◽  
Time Resolved ◽  
Single Molecule Level ◽  
Film Interpretation ◽  
Fluorescence Measurements ◽  
Naphthalene Diimide ◽  
Prompt And Delayed Fluorescence

Electron transfer (ET) is a key chemical reaction in nature and has been extensively studied in bulk systems, but remains challenging to investigate at the single-molecule level. A previously reported naphthalene diimide (NDI)-based system (Higginbotham et al., Chem. Commun. 2013, 49, 5061–5063) displays delayed fluorescence with good quantum yield (~0.5) and long-lived (nanoseconds) prompt and delayed fluorescence lifetimes, providing an opportunity to interrogate the underlying ET processes in single molecules. Time-resolved single-molecule fluorescence measurements enabled forward and reverse ET rate constants to be calculated for 45 individual molecules embedded in poly(methylmethacrylate) (PMMA) film. Interpretation of the results within the framework of Marcus–Hush theory for ET demonstrates that variation in both the electronic coupling and the driving force for ET is occurring from molecule to molecule within the PMMA film and over time for individual molecules.

Reversible Intramolecular Electron Transfer at the Single-Molecule Level

2003 ◽  
Vol 115 (35) ◽  
pp. 4341-4346 ◽  
Author(s):  
Roel Gronheid ◽  
Alina Stefan ◽  
Mircea Cotlet ◽  
Johan Hofkens ◽  
Jianqiag Qu ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Single Molecule ◽  
Intramolecular Electron Transfer ◽  
Single Molecule Level

scholarly journals Transition from stochastic events to deterministic ensemble average in electron transfer reactions revealed by single-molecule conductance measurement

2019 ◽  
Vol 116 (9) ◽  
pp. 3407-3412 ◽  
Author(s):  
Yueqi Li ◽  
Hui Wang ◽  
Zixiao Wang ◽  
Yanjun Qiao ◽  
Jens Ulstrup ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Single Molecule ◽  
Electron Transfer Reaction ◽  
Transfer Reactions ◽  
Ensemble Averaging ◽  
Electron Transfer Reactions ◽  
Single Molecule Level ◽  
Individual Reaction ◽  
Stochastic Events ◽  
The Individual

Electron transfer reactions can now be followed at the single-molecule level, but the connection between the microscopic and macroscopic data remains to be understood. By monitoring the conductance of a single molecule, we show that the individual electron transfer reaction events are stochastic and manifested as large conductance fluctuations. The fluctuation probability follows first-order kinetics with potential dependent rate constants described by the Butler–Volmer relation. Ensemble averaging of many individual reaction events leads to a deterministic dependence of the conductance on the external electrochemical potential that follows the Nernst equation. This study discloses a systematic transition from stochastic kinetics of individual reaction events to deterministic thermodynamics of ensemble averages and provides insights into electron transfer processes of small systems, consisting of a single molecule or a small number of molecules.

Electron transfer behaviour of biological macromolecules towards the single-molecule level

2003 ◽  
Vol 15 (18) ◽  
pp. S1873-S1890 ◽  
Author(s):  
Jingdong Zhang ◽  
Mikala Grubb ◽  
Allan G Hansen ◽  
Alexander M Kuznetsov ◽  
Anja Boisen ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Single Molecule ◽  
Biological Macromolecules ◽  
Single Molecule Level

Electron Transfer at the Single-Molecule Level in a Triphenylamine-Perylene Imide Molecule

ChemPhysChem ◽  
2005 ◽  
Vol 6 (5) ◽  
pp. 942-948 ◽  
Author(s):  
Toby D. M. Bell ◽  
Alina Stefan ◽  
Sadahiro Masuo ◽  
Tom Vosch ◽  
Marc Lor ◽  
...  
Keyword(s):  
Electron Transfer ◽  
Single Molecule ◽  
Single Molecule Level
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