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  4. Binding of Biotin to Streptavidin: A combined fluorescence correlation spectroscopy and time-resolved fluorescence study
 
research article

Binding of Biotin to Streptavidin: A combined fluorescence correlation spectroscopy and time-resolved fluorescence study

Stromqvist, J.
•
Nardo, L.
•
Broekmans, O.
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2011
European Physical Journal-Special Topics

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.

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Type
research article
DOI
10.1140/epjst/e2011-01513-1
Web of Science ID

WOS:000297814600012

Author(s)
Stromqvist, J.
Nardo, L.
Broekmans, O.
Kohn, J.
Lamperti, M.
Santamato, A.
Shalaby, M.
Sharma, G.
Di Trapani, P.
Bondani, M.
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Date Issued

2011

Published in
European Physical Journal-Special Topics
Volume

199

Start page

181

End page

194

Subjects

Stability

Editorial or Peer reviewed

REVIEWED

Written at

EPFL

EPFL units
IMT  
Available on Infoscience
June 12, 2012
Use this identifier to reference this record
https://infoscience.epfl.ch/handle/20.500.14299/81599
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