Bioenergetic shift and proteomic signature induced by lentiviral-transduction of GFP-based biosensors.

Autor:	Barakat S; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey., Çimen Ş; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey; Department of Nutrition and Dietetics, Institution of Health Sciences, Istanbul Medipol University, Istanbul, 34810, Turkey., Miri SM; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey; Molecular Biology, Genetics, and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, 34956, Turkey., Vatandaşlar E; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey., Yelkenci HE; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey., San Martín A; Centro de Estudios Científicos (CECs), 5110466, Valdivia, Chile; Facultad de Ciencias para el Cuidado de la Salud, Universidad San Sebastián, 5110773, Valdivia, Chile., Beker MÇ; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey; Department of Physiology, School of Medicine, Istanbul Medipol University, Istanbul, 34810, Turkey., Kök K; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey; Department of Biostatistics and Medical Informatics, International School of Medicine, Istanbul Medipol University, Istanbul, 34810, Turkey. Electronic address: guvanch@medipol.edu.tr., Öztürk G; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey; Department of Physiology, School of Medicine, Bolu Abant İzzet Baysal University, Bolu, 14030, Turkey. Electronic address: gurkan.ozturk@ibu.edu.tr., Eroglu E; Regenerative and Restorative Medicine Research Center (REMER), Research Institute for Health Sciences and Technologies (SABITA), Istanbul Medipol University, Istanbul, 34810, Turkey; Molecular Biology, Genetics, and Bioengineering Program, Faculty of Engineering and Natural Sciences, Sabanci University, Istanbul, 34956, Turkey. Electronic address: emrah.eroglu@medipol.edu.tr.
Jazyk:	angličtina
Zdroj:	Redox biology [Redox Biol] 2024 Dec; Vol. 78, pp. 103416. Date of Electronic Publication: 2024 Nov 02.
DOI:	10.1016/j.redox.2024.103416
Abstrakt:	Fluorescent proteins (FPs) stand as pivotal tools extensively employed across diverse biological research endeavors in various model systems. However, long-standing concerns surround their use due to the numerous side effects associated with their expression. Recent investigations have brought to light the significance of hydrogen peroxide (H 2 O 2 ) that is associated with the maturation process of green fluorescent protein (GFP) fluorophores. The structural and functional impairments associated with GFP expression are possibly linked to this amount of H 2 O 2 . In this study, we assess the impact of the GFP-based HyPer7 biosensor on cellular homeostasis and proteome changes, aiming to identify potential risks related to oxidative stress responses that potentially risks the application of such tools. Cells expressing genome-integrated HyPer7 demonstrated altered mitochondrial membrane potential (MMP), which was alleviated by the addition of antioxidants or culturing cells at physiological normoxia (5 kPa O 2 ). Additionally, HyPer7-expressing cells also exhibited significant impairment in mitochondrial oxidative respiration, suggesting broader mitochondrial dysfunction. Through untargeted proteomics analysis, we identified 26 proteins exhibiting differential expression in HyPer7-expressing cells compared to respective control cells. Functional annotation analysis showed that the list of the delineated proteins is associated with cellular responses to stress and the regulation of antioxidant mechanisms. Our findings underscore the significance of caution and validation in ensuring a thorough comprehension of cellular responses when using fluorescent protein-based tools, thereby enhancing the reliability of the results. Competing Interests: Declaration of competing interest None. (Copyright © 2024 The Authors. Published by Elsevier B.V. All rights reserved.)
Databáze:	MEDLINE
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