Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions

Dietz MS, Wehrheim SS, Harwardt M-LIE, Niemann H, Heilemann M (2019)
NANO LETTERS 19(11): 8245-8249.

Zeitschriftenaufsatz | Veröffentlicht | Englisch
 
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Autor*in
Dietz, Marina S.; Wehrheim, S. Sophia; Harwardt, Marie-Lena I. E.; Niemann, HartmutUniBi; Heilemann, Mike
Abstract / Bemerkung
Fluorescence methods are important tools in modern biology. Direct labeling of biomolecules with a fluorophore might, however, change interaction surfaces. Here, we introduce a competitive binding assay in combination with fluorescence correlation spectroscopy that reports binding affinities of both labeled and unlabeled biomolecules to their binding target. We investigated how fluorophore labels at different positions of a DNA oligonucleotide affect hybridization to a complementary oligonucleotide and found dissociation constants varying within 2 orders of magnitude. We next demonstrated that placing a fluorophore label at position Leu280 in the protein ligand internalin B does not alter the binding affinity to the MET receptor tyrosine kinase, compared to unlabeled internalin B. Our approach is simple to implement and can be applied to investigate the influence of fluorophore labels in a large variety of biomolecular interactions.
Stichworte
Binding affinity; competitive binding; fluorescence correlation; spectroscopy; fluorescence labeling; fluorescent probes; protein-protein; interactions
Erscheinungsjahr
2019
Zeitschriftentitel
NANO LETTERS
Band
19
Ausgabe
11
Seite(n)
8245-8249
ISSN
1530-6984
eISSN
1530-6992
Page URI
https://pub.uni-bielefeld.de/record/2939444

Zitieren

Dietz MS, Wehrheim SS, Harwardt M-LIE, Niemann H, Heilemann M. Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions. NANO LETTERS. 2019;19(11):8245-8249.
Dietz, M. S., Wehrheim, S. S., Harwardt, M. - L. I. E., Niemann, H., & Heilemann, M. (2019). Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions. NANO LETTERS, 19(11), 8245-8249. doi:10.1021/acs.nanolett.9b03736
Dietz, M. S., Wehrheim, S. S., Harwardt, M. - L. I. E., Niemann, H., and Heilemann, M. (2019). Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions. NANO LETTERS 19, 8245-8249.
Dietz, M.S., et al., 2019. Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions. NANO LETTERS, 19(11), p 8245-8249.
M.S. Dietz, et al., “Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions”, NANO LETTERS, vol. 19, 2019, pp. 8245-8249.
Dietz, M.S., Wehrheim, S.S., Harwardt, M.-L.I.E., Niemann, H., Heilemann, M.: Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions. NANO LETTERS. 19, 8245-8249 (2019).
Dietz, Marina S., Wehrheim, S. Sophia, Harwardt, Marie-Lena I. E., Niemann, Hartmut, and Heilemann, Mike. “Competitive Binding Study Revealing the Influence of Fluorophore Labels on Biomolecular Interactions”. NANO LETTERS 19.11 (2019): 8245-8249.

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