An enzyme like lysozyme is a major plyer in the human immune system and disease defense system. However, bacterial pathogens, in turn, have evolved resistance mechanisms to evade immune defense reaction. As consequences, super-resilient, drug-resistant pathogens are major issues in health care. The long-term objective of this proposal is to offer insight into the molecular interaction of different ligands including modified-substrates and inhibitors to lysozyme, which is a prerequisite for the development of novel antibacterial agents. Three integrated specific aims are proposed to reveal a precise molecular mechanism that controls dynamic protein-ligand interaction using a novel, electronic-type single-molecule approach developed by the PI. This approach interrogates enzyme dynamics and its involvement in ligand binding, catalysis, and inhibition at unprecedented time scales over extended periods of time measurements. With the invention, we will initially investigate the enzyme fluctuation and flexibility to determine their contribution to the ligand recognition and binding, as well as their roles to the enzyme catalysis and inhibition. Next, we will dissect the structural and molecular basis of enzyme-ligand interaction with various mimic antibacterial inhibitors, including modified-substrates and peptide-inhibitors. In addition, we plan to drive the fluctuation and examine the effects on enzyme activity. The research attempts to understand whether the driven-fluctuation can interfere with, or even control, enzyme activity and function, either by inhibiting or amplifying the native activity. The outcome of the proposed research will help establish a structural basis for the design of compounds that can suppress super-resilient, drug-resistant pathogens as well as provide new possibilities for diagnostic devices that bridge electronics and biochemical function.

Public Health Relevance

Due to the antibiotic overuse, super-resilient, drug-resistant bacterial pathogens that can work against innate immune proteins like lysozymes have become a growing healthcare problem. Relying on a new invention, the proposed work aims to interrogate and control the molecular interaction of protein- antibacterial inhibitors, which could advance our understanding of disease biology and suggest avenues toward the development of designing novel antibacterial drugs.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Academic Research Enhancement Awards (AREA) (R15)
Project #
1R15GM122063-01A1
Application #
9228830
Study Section
Biochemistry and Biophysics of Membranes Study Section (BBM)
Program Officer
Lewis, Catherine D
Project Start
2016-09-23
Project End
2019-08-31
Budget Start
2016-09-23
Budget End
2019-08-31
Support Year
1
Fiscal Year
2016
Total Cost
$423,892
Indirect Cost
$131,553
Name
North Dakota State University
Department
Physics
Type
Schools of Arts and Sciences
DUNS #
803882299
City
Fargo
State
ND
Country
United States
Zip Code
58108
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Froberg, James; Jayasooriya, Vidura; You, Seungyong et al. (2017) Quantitative measurements of dielectrophoresis in a nanoscale electrode array with an atomic force microscopy. Appl Phys Lett 110:203701