Dimer formation of organic fluorophores reports on biomolecular dynamics under denaturing conditions
| Autor: | Markus Sauer, Stefan Bollmann, Marc Löllmann, Soeren Doose |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
Protein Denaturation
Fluorophore Protein Conformation Dimer Analytical chemistry General Physics and Astronomy Fluorescence correlation spectroscopy Nucleic Acid Denaturation Photochemistry Motion chemistry.chemical_compound Biopolymers Oxazines Denaturation (biochemistry) Physical and Theoretical Chemistry Guanidine Fluorescent Dyes chemistry.chemical_classification Molecular Structure Biomolecule Fluorescence intermittency Fluorescence Kinetics Spectrometry Fluorescence chemistry Spectrophotometry Spectrophotometry Ultraviolet Dimerization Oligopeptides |
| DOI: | 10.1039/c1cp21111k |
| Popis: | Stacking interactions between organic fluorophores can cause formation of non-fluorescent H-dimers. Dimer formation and dissociation of two fluorophores site-specifically incorporated in a biomolecule result in fluorescence intermittency that can report on conformational dynamics. We characterize intramolecular dimerization of two oxazine fluorophores MR121 attached to an unstructured polypeptide. Formation of stable non-fluorescent complexes with nano- to microsecond lifetimes is a prerequisite for analysing the intermittent fluorescence emission by fluorescence correlation spectroscopy and extracting relaxation time constants on nano- to millisecond time scales. Destabilization of the generally very stable homodimers by chemical denaturation reduces the lifetime of H-dimers. We demonstrate that H-dimer formation of an oxazine fluorophore reports on end-to-end contact rates in unstructured glycine-serine polypeptides under denaturing conditions. |
| Databáze: | OpenAIRE |
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