title: The dityrosine cross-link as an intrinsic donor for assembling FRET pairs in the study of protein structure.
abstract: Dityrosine-Lucifer yellow (DT-LY) was used as the donor-acceptor pair in studies of the topography of juvenile hormone-binding protein (JHBP). The Förster distance, R(0)=30.5Å for DT-LY was determined. Separation distances (R) between DT and the fluorescent probes placed at the 219 and 224 position were 26 and 28Å and correspond to that found from X-ray analysis (23 and 24Å, respectively). Higher than expected efficiency of energy transfer between DT and LY probe placed in position 171 was observed, indicating that this probe is immobilized in the protein structure (κ(2)=3.25). Red-edge excitation shift (REES) analysis supported this assumption. Slight changes in Förster resonance energy transfer (FRET) efficiency were observed after incubating the labeled proteins with juvenile hormone III (JH III). This is the first report showing the application of DT fluorescence for the analysis of protein conformation.
authors.affiliation: Department of Biochemistry, Faculty of Chemistry, Wrocław University of Technology, Wybrzeże Wyspiańskiego 27, 50-370 Wrocław, Poland.
authors.forename: Dominika
authors.initials: D
authors.lastname: Bystranowska
authors.forename: Beata
authors.initials: B
authors.lastname: Siejda
authors.forename: Andrzej
authors.initials: A
authors.lastname: Ożyhar
authors.forename: Marian
authors.initials: M
authors.lastname: Kochman
mesh_terms: D003432:Cross-Linking Reagents / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer; D005456:Fluorescent Dyes / Q000737:chemistry*; D007546:Isoquinolines / Q000737:chemistry; D007605:Juvenile Hormones / Q000737:chemistry* / Q000235:genetics; D008958:Models, Molecular; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D012717:Sesquiterpenes / Q000737:chemistry; D014443:Tyrosine / Q000031:analogs & derivatives* / Q000737:chemistry
chemical_list: D003432:Cross-Linking Reagents; D005456:Fluorescent Dyes; D007546:Isoquinolines; D007605:Juvenile Hormones; D011994:Recombinant Proteins; D012717:Sesquiterpenes; D014443:Tyrosine; C017475:lucifer yellow; C036585:juvenile hormone III; C007543:dityrosine