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Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions

Cellular conformation of reduced pyridine nucleotides NADH and NADPH sensed using autofluorescence spectroscopy is presented as a real-time metabolic indicator under pressurized conditions. The approach provides information on the role of pressure in energy metabolism and antioxidant defense with ap...

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Autores principales: Heidelman, Martin, Dhakal, Bibek, Gikunda, Millicent, Silva, Kalinga Pavan Thushara, Risal, Laxmi, Rodriguez, Andrew I., Abe, Fumiyoshi, Urayama, Paul
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402201/
https://www.ncbi.nlm.nih.gov/pubmed/34443607
http://dx.doi.org/10.3390/molecules26165020
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author Heidelman, Martin
Dhakal, Bibek
Gikunda, Millicent
Silva, Kalinga Pavan Thushara
Risal, Laxmi
Rodriguez, Andrew I.
Abe, Fumiyoshi
Urayama, Paul
author_facet Heidelman, Martin
Dhakal, Bibek
Gikunda, Millicent
Silva, Kalinga Pavan Thushara
Risal, Laxmi
Rodriguez, Andrew I.
Abe, Fumiyoshi
Urayama, Paul
author_sort Heidelman, Martin
collection PubMed
description Cellular conformation of reduced pyridine nucleotides NADH and NADPH sensed using autofluorescence spectroscopy is presented as a real-time metabolic indicator under pressurized conditions. The approach provides information on the role of pressure in energy metabolism and antioxidant defense with applications in agriculture and food technologies. Here, we use spectral phasor analysis on UV-excited autofluorescence from Saccharomyces cerevisiae (baker’s yeast) to assess the involvement of one or multiple NADH- or NADPH-linked pathways based on the presence of two-component spectral behavior during a metabolic response. To demonstrate metabolic monitoring under pressure, we first present the autofluorescence response to cyanide (a respiratory inhibitor) at 32 MPa. Although ambient and high-pressure responses remain similar, pressure itself also induces a response that is consistent with a change in cellular redox state and ROS production. Next, as an example of an autofluorescence response altered by pressurization, we investigate the response to ethanol at ambient, 12 MPa, and 30 MPa pressure. Ethanol (another respiratory inhibitor) and cyanide induce similar responses at ambient pressure. The onset of non-two-component spectral behavior upon pressurization suggests a change in the mechanism of ethanol action. Overall, results point to new avenues of investigation in piezophysiology by providing a way of visualizing metabolism and mitochondrial function under pressurized conditions.
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spelling pubmed-84022012021-08-29 Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions Heidelman, Martin Dhakal, Bibek Gikunda, Millicent Silva, Kalinga Pavan Thushara Risal, Laxmi Rodriguez, Andrew I. Abe, Fumiyoshi Urayama, Paul Molecules Article Cellular conformation of reduced pyridine nucleotides NADH and NADPH sensed using autofluorescence spectroscopy is presented as a real-time metabolic indicator under pressurized conditions. The approach provides information on the role of pressure in energy metabolism and antioxidant defense with applications in agriculture and food technologies. Here, we use spectral phasor analysis on UV-excited autofluorescence from Saccharomyces cerevisiae (baker’s yeast) to assess the involvement of one or multiple NADH- or NADPH-linked pathways based on the presence of two-component spectral behavior during a metabolic response. To demonstrate metabolic monitoring under pressure, we first present the autofluorescence response to cyanide (a respiratory inhibitor) at 32 MPa. Although ambient and high-pressure responses remain similar, pressure itself also induces a response that is consistent with a change in cellular redox state and ROS production. Next, as an example of an autofluorescence response altered by pressurization, we investigate the response to ethanol at ambient, 12 MPa, and 30 MPa pressure. Ethanol (another respiratory inhibitor) and cyanide induce similar responses at ambient pressure. The onset of non-two-component spectral behavior upon pressurization suggests a change in the mechanism of ethanol action. Overall, results point to new avenues of investigation in piezophysiology by providing a way of visualizing metabolism and mitochondrial function under pressurized conditions. MDPI 2021-08-19 /pmc/articles/PMC8402201/ /pubmed/34443607 http://dx.doi.org/10.3390/molecules26165020 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Heidelman, Martin
Dhakal, Bibek
Gikunda, Millicent
Silva, Kalinga Pavan Thushara
Risal, Laxmi
Rodriguez, Andrew I.
Abe, Fumiyoshi
Urayama, Paul
Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions
title Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions
title_full Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions
title_fullStr Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions
title_full_unstemmed Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions
title_short Cellular NADH and NADPH Conformation as a Real-Time Fluorescence-Based Metabolic Indicator under Pressurized Conditions
title_sort cellular nadh and nadph conformation as a real-time fluorescence-based metabolic indicator under pressurized conditions
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8402201/
https://www.ncbi.nlm.nih.gov/pubmed/34443607
http://dx.doi.org/10.3390/molecules26165020
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