Abstract
The Biotin-Streptavidin complex is a widely studied system in biology and biophysics, because of its extremely strong non-covalent binding affinity. The latter is often exploited to link molecules to substrates or to one another. However, the details of the Biotin-Streptavidin binding have not been fully elucidated so far. Particularly, the role of cooperative effects in enhancing the binding affinity has not been clarified. Our long-term aim is to investigate this point by implementing two complementary approaches, fluorescence correlation spectroscopy and time-correlated single-photon counting. As both methods rely on the analysis of fluorescence signals, biotin labeled with Atto-550-dye was used. In this work, in order to get a first overview of the system, we analyzed solutions in three paradigmatic ranges of Biotin-to-Streptavidin concentration ratio. Fluorescence correlation spectroscopy measurements allowed us to extract diffusion times of free biotin and of biotin-Streptavidin complexes, and also to gain information about the dynamics of the intersystem crossing between the first excited triplet and the first excited singlet states. Time-correlated single-photon counting made it possible to derive the lifetimes of the different species in solution, as well as to deduce relevant information about the relative abundance of Streptavidin-complexed and free Biotin.
Similar content being viewed by others
References
A. Holmberg, A. Blomstergren, O. Nord, M. Lukacs, J. Lundeberg, M. Uhlen, Electrophoresis 26, 501 (2005)
K. Svoboda, S.M. Block, Annu. Rev. Biophys. Biomol. Struct. 23, 247 (1994)
S.B. Smith, Y. Cui, C. Bustamante, Science 271, 795 (1996)
H. Kim, I.-H. Cho, J.H. Park, S. Kim, S.-H. Paek, J. Noh, H. Lee, Colloids and Surfaces A: Physicochem. Eng. Aspects 313–314, 541 (2007)
M. Lenhert, M. Gorbahn, C. Rosin, M. Klein, I. Köper, B. Al-Nawas, W. Knoll, M. Veith, Langmuir 27, 7743 (2011)
M. Gonzales, C.E. Argaraña, G.D. Fidelio, Biomol. Eng. 16, 67 (1999)
T. Sano, C.R. Cantor, J. Biol. Chem. 265, 3369 (1990)
M.L. Jones, G.P. Kurzban, Biochem. 34, 11750 (1995)
A. Loosli, U.E. Rusbandi, J. Gradinaru, K. Bernauer, C.W. Schlaepfer, M. Meyer, S. Mazurek, M. Novic, T.R. Ward, Inorg. Chem. 45, 660 (2006)
R. Rigler, U. Mets, J. Widengren, P. Kask, Eur. Biophys. J. 22, 169 (1993)
P. Schwille, Cell Biochem. Biophys. 34, 283 (2001)
M. Goesch, R. Rigler, Adv. Drug Deliv. Rev. 57, 169 (2005)
ATTO-TEC 2011 Online Catalogue: http://www.atto-tec.com/index.php?id=54∖ no_cache=1∖L=1
A. Andreoni, M. Bondani, L. Nardo, Mol. Cell. Probes 23, 119 (2009)
W. Becker, The bh TCSPC Handbook – Third Edition (Becker & Hickl, GmbH, 2008), p. 401
S. Freitag, I. Le Trang, L. Klumb, P.S. Stayton, R.E. Stenkamp, Prot. Sci. 6, 1157 (1997)
A. Einstein, Ann. der Physik 17, 549 (1905)
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Strömqvist, J., Nardo, L., Broekmans, O. et al. Binding of Biotin to Streptavidin: A combined fluorescence correlation spectroscopy and time-resolved fluorescence study. Eur. Phys. J. Spec. Top. 199, 181–194 (2011). https://doi.org/10.1140/epjst/e2011-01513-1
Received:
Revised:
Published:
Issue Date:
DOI: https://doi.org/10.1140/epjst/e2011-01513-1