scholarly journals Single-Molecule Imaging of anin Vitro-Evolved RNA Aptamer Reveals Homogeneous Ligand Binding Kinetics

2009 ◽  
Vol 131 (29) ◽  
pp. 9866-9867 ◽  
Author(s):  
Mark P. Elenko ◽  
Jack W. Szostak ◽  
Antoine M. van Oijen
2015 ◽  
Vol 112 (46) ◽  
pp. 14230-14235 ◽  
Author(s):  
Sándor Volkán-Kacsó ◽  
Rudolph A. Marcus

A theoretical model of elastically coupled reactions is proposed for single molecule imaging and rotor manipulation experiments on F1-ATPase. Stalling experiments are considered in which rates of individual ligand binding, ligand release, and chemical reaction steps have an exponential dependence on rotor angle. These data are treated in terms of the effect of thermodynamic driving forces on reaction rates, and lead to equations relating rate constants and free energies to the stalling angle. These relations, in turn, are modeled using a formalism originally developed to treat electron and other transfer reactions. During stalling the free energy profile of the enzymatic steps is altered by a work term due to elastic structural twisting. Using biochemical and single molecule data, the dependence of the rate constant and equilibrium constant on the stall angle, as well as the Børnsted slope are predicted and compared with experiment. Reasonable agreement is found with stalling experiments for ATP and GTP binding. The model can be applied to other torque-generating steps of reversible ligand binding, such as ADP and Pi release, when sufficient data become available.


2020 ◽  
Author(s):  
Thilini Perera ◽  
Hirushi Gunasekara ◽  
Ying S. Hu

Single-molecule imaging has provided new insights on weak transient biomolecular interactions with micromolar to millimolar affinity. However, the limited duration of observation has hindered the study of strong and reversible interactions with sub-nanomolar affinity. We report single-molecule interaction microscopy (SMIM), which combines point accumulation for imaging in nanoscale topography (PAINT) with extended imaging durations that enables the study of antibody binding kinetics in the cellular environment. SMIM revealed heterogeneous binding kinetics and the effect of concentration and antibody valency on the association and dissociation rates on antibody-antigen interactions in their cellular environments. We thereby demonstrate SMIM as a versatile single-molecule technique for studying strong, transient biomolecular interactions.


2009 ◽  
Vol 81 (1) ◽  
pp. 336-342 ◽  
Author(s):  
Joshua R. Wayment ◽  
Joel M. Harris

2009 ◽  
Vol 131 (14) ◽  
pp. 5052-5053 ◽  
Author(s):  
Michael P. Latham ◽  
Grant R. Zimmermann ◽  
Arthur Pardi

NanoImpact ◽  
2017 ◽  
Vol 6 ◽  
pp. 90-98 ◽  
Author(s):  
Navid B. Saleh ◽  
Dipesh Das ◽  
Jaime Plazas-Tuttle ◽  
Darwin Yang ◽  
Jackson Travis Del Bonis-O'Donnell ◽  
...  

2016 ◽  
Vol 110 (3) ◽  
pp. 238a
Author(s):  
Max Kushner ◽  
Alexander Van Slyke ◽  
Fabio Rinaldi ◽  
Avtar Singh ◽  
John Lis ◽  
...  

Sign in / Sign up

Export Citation Format

Share Document