Skip to main content
ARS Home » Plains Area » Houston, Texas » Children's Nutrition Research Center » Research » Publications at this Location » Publication #415290
Research Project: Nutritional Role of Phytochemicals

Location: Children's Nutrition Research Center

Title: Development of a novel amplifiable system to quantify hydrogen peroxide in living cells

Author
item WANG, LINGFEI - Baylor College Of Medicine
item LIN, HANFENG - Baylor College Of Medicine
item YANG, BIN - Baylor College Of Medicine
item JIANG, XIQIAN - Baylor College Of Medicine
item CHEN, JIANWEI - Baylor College Of Medicine
item CHOWDHURY, SANDIPAN - Baylor College Of Medicine
item CHENG, NINGHUI - Children'S Nutrition Research Center (CNRC)
item Nakata, Paul
item LONARD, DAVID - Baylor College Of Medicine
item WANG, MENG - Baylor College Of Medicine
item WANG, JIN - Baylor College Of Medicine

Submitted to: Journal of the American Chemical Society
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/16/2024
Publication Date: 7/30/2024
Citation: Wang, L., Lin, H., Yang, B., Jiang, X., Chen, J., Chowdhury, S.R., Cheng, N., Nakata, P.A., Lonard, D.M., Wang, M.C., Wang, J. 2024. Development of a novel amplifiable system to quantify hydrogen peroxide in living cells. Journal of the American Chemical Society. 146:22396-22404. https://doi.org/10.1021/jacs.4c05366.
DOI: https://doi.org/10.1021/jacs.4c05366

Interpretive Summary: Living cells contain many chemicals that are important components in biological processes. Accurate measurements of these chemicals are often difficult because they are present in such low amounts within the cell. In this study we report the establishment of a new method to accurately measure chemicals that are present within the cell in small amounts. This method uses a biologically derived system, consisting of a receptor and enzyme, to amplify the measurement signal so that it can be accurately determined. We show the utility of this method by measuring the amount of the chemical, hydrogen peroxide, inside living cells. It is anticipated that this method can be adapted for the measurement of other chemicals found in low amounts within cells. It is our hope that this measurement system will find wide use in efforts to understand the importance of low abundance chemicals in biological processes.

Technical Abstract: Although many redox signaling molecules are present at low concentrations, typically ranging from micromolar to sub-micromolar levels, they often play essential roles in a wide range of biological pathways and disease mechanisms. However, accurately measuring low abundant analytes has been a significant challenge due to the lack of sensitivity and quantitative capability of existing measurement methods. In this study, we introduced a novel chemically induced amplifiable system for quantifying low-abundance redox signaling molecules in living cells. We utilized H2O2 as a proof-of concept analyte and developed a probe that quantifies cellular peroxide levels by combining the NanoBiT system with androgen receptor (AR) dimerization as a reporting mechanism. Our system demonstrated a highly sensitive response to cellular peroxide changes induced both endogenously and exogenously. Furthermore, the system can be adapted for the quantification of other signaling molecules if provided with suitable probing chemistry.