| Author |
Ameen, Abid Chang, Te-Wei Das, Aditi Logan Liu, Gang Plucinski, Lisa Hsiao, Austin Ranjan Gartia, Manas Arnold, William R. |
| Description |
Country affiliation: United States Author Affiliation: Plucinski L ( Department of Electrical and Computer Engineering, Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States.); Ranjan Gartia M ( Department of Electrical and Computer Engineering, Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States.); Arnold WR ( Department of Biochemistry, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States.); Ameen A ( Department of Materials Science and Engineering, Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States.); Chang TW ( Department of Electrical and Computer Engineering, Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States.); Hsiao A ( Department of Bioengineering, Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States.); Logan Liu G ( Department of Electrical and Computer Engineering, Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States. Electronic address: loganliu@illinois.edu.); Das A ( Department of Biochemistry, University of Illinois at Urbana-Champaign, 208 North Wright Street, Urbana, IL 61801, United States) |
| Abstract |
Cytochrome P450s are the primary enzymes involved in phase I drug metabolism. They are an important target for early drug discovery research. However, high-throughput drug screening of P450s is limited by poor protein stability and lack of consistent measurement of binding events. Here we present the detection of substrate binding to cytochrome P450-2J2 (CYP2J2), the predominant P450 in the human heart, using a combination of Nanodisc technology and a nanohole plasmonic sensor called nanoplasmonic Lycurgus cup array (nanoLCA). The Nanodisc, a nanoscale membrane bilayer disc, is used to stabilize the protein on the metallic plasmonic surface. Absorption spectroscopy of seven different substrates binding to CYP2J2 in solution showed that they are all type I, resulting in shifting of the protein bands to lower wavelengths (blue shift). Detection on the nanoLCA sensor also showed spectral blue shifts of CYP2J2 following substrate binding. Finite Difference Time Domain (FDTD) electromagnetic simulation suggested that the blue shift on the nanoLCA is because of the hybridization of plasmon polariton Bloch wave and the electronic resonance of the heme group of CYP2J2. We found the plasmonic properties of the nanoLCA sensor to be highly reproducible, which allowed comparisons among the different substrates at different concentrations. Further, due to the unique spectral properties of the nanoLCA sensor, including the transmission of a single color, we were able to perform colorimetric detection of the binding events. These results indicate that a resonance plasmonic sensing mechanism can be used to distinguish between different substrates of the same binding type at different concentrations binding to P450s and that the nanoLCA sensor has the potential to provide consistent high-throughput measurements of this system. |
