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The Use of ProteoTuner Technology to Study Nuclear Factor κB Activation by Heavy Ions

Chishti, Arif Ali and Baumstark-Khan, Christa and Nisar, Hasan and Hu, Yueyuan and Konda, Bikash and Henschenmacher, Bernd and Spitta, Luis F. and Schmitz, Claudia and Feles, Sebastian and Hellweg, Christine E. (2021) The Use of ProteoTuner Technology to Study Nuclear Factor κB Activation by Heavy Ions. International Journal of Molecular Sciences, 22 (24), p. 13530. Multidisciplinary Digital Publishing Institute (MDPI). doi: 10.3390/ijms222413530. ISSN 1661-6596.

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Official URL: https://www.mdpi.com/1422-0067/22/24/13530

Abstract

Nuclear factor κB (NF-κB) activation might be central to heavy ion-induced detrimental processes such as cancer promotion and progression and sustained inflammatory responses. A sensitive detection system is crucial to better understand its involvement in these processes. Therefore, a DD-tdTomato fluorescent protein-based reporter system was previously constructed with human embryonic kidney (HEK) cells expressing DD-tdTomato as a reporter under the control of a promoter containing NF-κB binding sites (HEK-pNFκB-DD-tdTomato-C8). Using this reporter cell line, NF-κB activation after exposure to different energetic heavy ions (¹⁶O, 95 MeV/n, linear energy transfer—LET 51 keV/µm; ¹²C, 95 MeV/n, LET 73 keV/μm; ³⁶Ar, 95 MeV/n, LET 272 keV/µm) was quantified considering the dose and number of heavy ions hits per cell nucleus that double NF-κB-dependent DD-tdTomato expression. Approximately 44 hits of ¹⁶O ions and ≈45 hits of ¹²C ions per cell nucleus were required to double the NF-κB-dependent DD-tdTomato expression, whereas only ≈3 hits of ³⁶Ar ions were sufficient. In the presence of Shield-1, a synthetic molecule that stabilizes DD-tdTomato, even a single particle hit of ³⁶Ar ions doubled NF-κB-dependent DD-tdTomato expression. In conclusion, stabilization of the reporter protein can increase the sensitivity for NF-κB activation detection by a factor of three, allowing the detection of single particle hits’ effects.

Item URL in elib:https://elib.dlr.de/147989/
Document Type:Article
Additional Information:This article belongs to the Special Issue Biological Effects of Low Dose and Low Dose-Rate Ionizing Radiation Exposure
Title:The Use of ProteoTuner Technology to Study Nuclear Factor κB Activation by Heavy Ions
Authors:
AuthorsInstitution or Email of AuthorsAuthor's ORCID iD
Chishti, Arif AliRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany and Department of Biochemistry and Molecular Biology, School of Medicine, Shenzhen University, China; arifalichishti (at) szu.edu.cnUNSPECIFIED
Baumstark-Khan, ChristaRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany; Christa.Baumstark-Khan (at) dlr.dehttps://orcid.org/0000-0002-9329-0128
Nisar, HasanRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany and Department of Medical Sciences, Pakistan Institute of Engineering and Applied Sciences (PIEAS), Lehtrar Road, Nilore, Islamabad, PakistanUNSPECIFIED
Hu, YueyuanRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany and Institute of Cardiovascular Immunology, University Hospital Bonn, D-53127 Bonn, GermanyUNSPECIFIED
Konda, BikashRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany; bikash.konda (at) dlr.deUNSPECIFIED
Henschenmacher, BerndRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany and Competence Center for Electromagnetic Fields (KEMF), Federal Office for Radiation Protection, Oberschleißheim, GermanyUNSPECIFIED
Spitta, Luis F.Radiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, GermanyUNSPECIFIED
Schmitz, ClaudiaRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, GermanyUNSPECIFIED
Feles, SebastianRadiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany and Gravitational Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, GermanyUNSPECIFIED
Hellweg, Christine E.Radiation Biology Department, Institute of Aerospace Medicine, German Aerospace Center (DLR), Cologne, Germany; christine.hellweg (at) dlr.dehttps://orcid.org/0000-0002-2223-3580
Date:16 December 2021
Journal or Publication Title:International Journal of Molecular Sciences
Refereed publication:Yes
Open Access:Yes
Gold Open Access:Yes
In SCOPUS:Yes
In ISI Web of Science:Yes
Volume:22
DOI:10.3390/ijms222413530
Page Range:p. 13530
Publisher:Multidisciplinary Digital Publishing Institute (MDPI)
ISSN:1661-6596
Status:Published
Keywords:nuclear factor κB; reporter system; ProteoTuner system; heavy ions; X-rays; tdTomato; DD-tdTomato; fluorescent protein; galactic cosmic rays; space missions
HGF - Research field:Aeronautics, Space and Transport
HGF - Program:Space
HGF - Program Themes:Research under Space Conditions
DLR - Research area:Raumfahrt
DLR - Program:R FR - Research under Space Conditions
DLR - Research theme (Project):R - Project ISS LIFE 2.0
Location: Köln-Porz
Institutes and Institutions:Institute of Aerospace Medicine > Radiation Biology
Institute of Aerospace Medicine > Gravitational Biology
Deposited By: Kopp, Kerstin
Deposited On:10 Jan 2022 08:22
Last Modified:10 Jan 2022 08:22

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