Enzymatic Single-Molecule Spectroelectrochemistry by Nanoscale Confinement and Nanometer Scale Super Resolution Fluorescence Imaging
EMSL Project ID
50719
Abstract
We propose a new multi-disciplinary approach via single-molecule spectroscopy to study enzyme-catalyzed single-molecule electron transfer reaction and image the redox kinetics of enzymes in nanometer scale resolution under controlled electrochemical potential. We will chemically modify the enzyme environment to confine the set of conformations available to the enzyme and electron transfer mediator, if needed, to measure the effects of reduced conformational freedom on electron transfer reaction rates and study the enzymatic reaction mechanism in depth at single-molecule level. Selective, site-directed tagging and tethering of the same target enzymes leading to new understanding of the relationships between rate constant fluctuations (dynamic disorder) and the degrees of freedom of the active site and surrounding enzyme structures. Cyclic voltammetry will be coupled to single-molecule spectroscopy to measure the reaction rates as a function of chemical potential. The results with enzymes involved in electron transfer reactions will enable a deeper understanding of the fundamental factors controlling the rapid conformational changes affecting enzymatic activity. The knowledge developed using these techniques will enable us to understand the fundamental enzymatic electron transfer reactions in depth and details as well as the metabolism of microbial communities and their impact to the environment.
Project Details
Start Date
2019-05-01
End Date
2021-09-30
Status
Closed
Released Data Link
Team
Principal Investigator
Team Members
Related Publications
Dehong Hu, Chenghong Lei. 2021. "High Throughput Mapping of Single Molecules’ Redox Potentials on Electrode." Analytical Chemistry 93 (25):8864-8871. 10.1021/acs.analchem.1c00984
Ming Chen, Dehong Hu, Chenghong Lei, Richard C. Zangar. 2021. "Colloidal immobilized protein based on stable colloid of TiO nanoparticles at neutral pH for protein microarray." Colloid and Interface Science Communications 43:100440. 10.1016/j.colcom.2021.100440