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To monitor conformational changes of individual membrane transporters in liposomes in real time, we attach two fluorophores to selected domains of a protein. Sequential distance changes between the dyes are recorded and analyzed by Forster…

定量方法 · 定量生物学 2015-06-11 Bertram Su , Monika G. Dueser , Nawid Zarrabi , Thomas Heitkamp , Ilka Starke , Michael Börsch

Observation times of freely diffusing single molecules in solution are limited by the photophysics of the attached fluorescence markers and by a small observation volume in the femtolitre range that is required for a sufficient…

Trapping nanoscopic objects to observe their dynamic behaviour for extended periods of time is an ongoing quest. Particularly, sub-100nm transparent objects are hard to catch and most techniques rely on immobilisation or transient diffusion…

定量方法 · 定量生物学 2017-01-13 Maria Dienerowitz , Thomas Heitkamp , Thomas Gottschall , Jens Limpert , Michael Borsch

FoF1-ATP synthase is the essential membrane enzyme maintaining the cellular level of adenosine triphosphate (ATP) and comprises two rotary motors. We measure subunit rotation in FoF1-ATP synthase by intramolecular Foerster resonance energy…

定量方法 · 定量生物学 2015-05-27 Torsten Rendler , Marc Renz , Eva Hammann , Stefan Ernst , Nawid Zarrabi , Michael Boersch

Single-molecule F\"orster resonance energy transfer (smFRET) of molecular motors provides transformative insights into their dynamics and conformational changes both at high temporal and spatial resolution simultaneously. However, a key…

FoF1-ATP synthase is the membrane protein catalyzing the synthesis of the 'biological energy currency' adenosine triphosphate (ATP). The enzyme uses internal subunit rotation for the mechanochemical conversion of a proton motive force to…

生物大分子 · 定量生物学 2015-06-15 Thomas Heitkamp , Hendrik Sielaff , Anja Korn , Marc Renz , Nawid Zarrabi , Michael Boersch

FoF1-ATP synthase catalyzes the synthesis of adenosine triphosphate (ATP). The F1 portion can be stripped from the membrane-embedded Fo portion of the enzyme. F1 acts as an ATP hydrolyzing enzyme, and ATP hydrolysis is associated with…

生物大分子 · 定量生物学 2018-02-14 Hendrik Sielaff , Thomas Heitkamp , Andrea Zappe , Nawid Zarrabi , Michael Boersch

Anti-Brownian traps confine single particles in free solution by closed-loop feedback forces that directly counteract Brownian motion. The extended-duration measurement of trapped objects allows detailed characterization of photophysical…

仪器与探测器 · 物理学 2023-05-15 Allison H. Squires , Abhijit A. Lavania , Peter D. Dahlberg , W. E. Moerner

Conformational changes of single proteins are monitored in real time by Forster-type resonance energy transfer, FRET. Two different fluorophores have to be attached to those protein domains, which move during function. The distance between…

生物物理 · 物理学 2009-11-13 N. Zarrabi , T. Heitkamp , J. C. Greie , M. Boersch

FoF1-ATP synthases are ubiquitous membrane-bound, rotary motor enzymes that can catalyze ATP synthesis and hydrolysis. Their enzyme kinetics are controlled by internal subunit rotation, by substrate and product concentrations, by mechanical…

生物大分子 · 定量生物学 2021-06-29 Thomas Heitkamp , Michael Börsch

Multiplexed, real-time fluorescence detection at the single-molecule level is highly desirable to reveal the stoichiometry, dynamics, and interactions of individual molecular species within complex systems. However, traditionally…

F1-ATPase is the soluble portion of the membrane-embedded enzyme FoF1-ATP synthase that catalyzes the production of adenosine triphosphate in eukaryotic and eubacterial cells. In reverse, the F1 part can also hydrolyze ATP quickly at three…

生物大分子 · 定量生物学 2015-06-18 Samuel D. Bockenhauer , Thomas M. Duncan , W. E. Moerner , Michael Boersch

Enzymes are cellular protein machines using a variety of conformational changes to power fast biochemical catalysis. Our goal is to exploit the single-spin properties of the luminescent NV (nitrogen-vacancy) center in nanodiamonds to reveal…

定量方法 · 定量生物学 2024-01-25 Ivan Perez , Anke Krueger , Joerg Wrachtrup , Fedor Jelezko , Michael Börsch

Plasmonic nanostructures can overcome Abbe's diffraction limit to generate strong gradient fields, enabling efficient optical trapping of nano-sized particles. However, it remains challenging to achieve stable trapping with low incident…

光学 · 物理学 2020-06-24 Domna G. Kotsifaki , Viet Giang Truong , Sile Nic Chormaic

Diffusion processes in biological membranes are of interest to understand the macromolecular organisation and function of several molecules. Fluorescence Recovery After Photobleaching (FRAP) has been widely used as a method to analyse this…

统计力学 · 物理学 2007-05-23 C. Favard , N. Olivi-Tran , J. -L. Meunier

Light scattering inhibits high-resolution optical imaging, manipulation and therapy deep inside biological tissue by preventing focusing. To form deep foci, wavefront-shaping and time-reversal techniques that break the optical diffusion…

光学 · 物理学 2015-06-04 Cheng Ma , Fengbo Zhou , Yan Liu , Lihong V. Wang

ATP synthases utilize a proton motive force to synthesize ATP. In reverse, these membrane-embedded enzymes can also hydrolyze ATP to pump protons over the membrane. To prevent wasteful ATP hydrolysis, distinct control mechanisms exist for…

生物大分子 · 定量生物学 2018-01-16 Hendrik Sielaff , Dhirendra Singh , Gerhard Grueber , Michael Börsch

Fluorescence resonance energy transfer (FRET) from individual, dye-labeled RNA molecules confined in freely-diffusing attoliter-volume aqueous droplets is carefully compared to FRET from unconfined RNA in solution. The use of…

生物物理 · 物理学 2015-05-21 Sheema Rahmanseresht , Peker Milas , Kieran P. Ramos , Ben D. Gamari , Lori S. Goldner

We present a Ramsey-type atom interferometer operating with an optically trapped sample of 10^6 Bose-condensed Rb-87 atoms. The optical trap allows us to couple the |F =1, mF =0>\rightarrow |F =2, mF =0> clock states using a single photon…

Adenosine triphosphate (ATP) is the universal chemical energy currency for cellular activities provided mainly by the membrane enzyme FoF1-ATP synthase in bacteria, chloroplasts and mitochondria. Synthesis of ATP is accompanied by subunit…

生物大分子 · 定量生物学 2016-08-03 Thomas Heitkamp , Gabriele Deckers-Hebestreit , Michael Börsch
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