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Extending Bio-SAXS measurements of Single-Stranded DNA-Binding Proteins: Radiation Protection of G5P by Cosolutes

  • Small-angle X-ray scattering (SAXS) can be used for structural de- termination of biological macromolecules and polymers in their na- tive states. To improve the reliability of such experiments, the re- duction of radiation damage occurring from exposure to X-rays is needed.One method, is the use of scavenger molecules that protect macromolecules against radicals produced by radiation exposure.In this study we investigate the feasibility to apply the compatible solute, osmolyte and radiation protector Ectoine (THP(B)) as a scavenger throughout SAXS measurements of single-stranded DNA-binding protein Gene-V Protein (G5P/GVP). Therefore we monitor the radiation induced changes of G5P during bio-SAXS. The resulting microscopic energy-damage relation was determined by particle scattering simu- lations with TOPAS/Geant4. The results are interpreted in terms of radical scavenging as well as post-irradiation effects, related to preferential-exclusion from the protein surface. Thus, EctoineSmall-angle X-ray scattering (SAXS) can be used for structural de- termination of biological macromolecules and polymers in their na- tive states. To improve the reliability of such experiments, the re- duction of radiation damage occurring from exposure to X-rays is needed.One method, is the use of scavenger molecules that protect macromolecules against radicals produced by radiation exposure.In this study we investigate the feasibility to apply the compatible solute, osmolyte and radiation protector Ectoine (THP(B)) as a scavenger throughout SAXS measurements of single-stranded DNA-binding protein Gene-V Protein (G5P/GVP). Therefore we monitor the radiation induced changes of G5P during bio-SAXS. The resulting microscopic energy-damage relation was determined by particle scattering simu- lations with TOPAS/Geant4. The results are interpreted in terms of radical scavenging as well as post-irradiation effects, related to preferential-exclusion from the protein surface. Thus, Ectoine provides an non-disturbing way to improve structure-determination of proteins via bio-SAXS in future studies.zeige mehrzeige weniger

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Metadaten
Autor*innen:Marc Benjamin HahnORCiD
Koautor*innen:Dorothea C. Hallier, Glen Jacob Smales, H. Seitz
Dokumenttyp:Posterpräsentation
Veröffentlichungsform:Präsentation
Sprache:Englisch
Jahr der Erstveröffentlichung:2023
Organisationseinheit der BAM:6 Materialchemie
6 Materialchemie / 6.6 Physik und chemische Analytik der Polymere
DDC-Klassifikation:Naturwissenschaften und Mathematik / Chemie / Analytische Chemie
Technik, Medizin, angewandte Wissenschaften / Ingenieurwissenschaften / Ingenieurwissenschaften und zugeordnete Tätigkeiten
Freie Schlagwörter:Ectoin
Bio-SAXS; BioSAXS; Compatible solute; Cosolute; DNA; Dosimetry; Ectoin; Ectoine; G5P; GVP; Geant4; Geant4-DNA; Gene five protein; Hydroxyectoine; Ionizing radiation damage; LEE; McSAS3; Median lethal energy deposit; Microdosimetry; Monte-Carlo simulations; OH Radical; OH radical scavenger; Osmolyte; Particle scattering simulations; Protein; Protein unfolding; Proteins; ROS; Radiation damage; Radical Scavenger; Radical scavenger; SAXS; Single-stranded DNA-binding proteins; Small-angle xray scattering; Topas; Topas-MC; Topas-nBio; X-ray scattering; ssDNA
Themenfelder/Aktivitätsfelder der BAM:Chemie und Prozesstechnik
Material
Material / Nano
Veranstaltung:MultiChem Conference 2023
Veranstaltungsort:Prague, Czech Republic
Beginndatum der Veranstaltung:26.04.2023
Enddatum der Veranstaltung:28.04.2023
Zugehöriger Identifikator:https://doi.org/10.1039/D2CP05053F
Verfügbarkeit des Dokuments:Datei im Netzwerk der BAM verfügbar ("Closed Access")
Datum der Freischaltung:04.05.2023
Referierte Publikation:Nein
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