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Journal of Fluorescence

, Volume 18, Issue 3–4, pp 601–610 | Cite as

Translational and Rotational Motions of Albumin Sensed by a Non-Covalent Associated Porphyrin Under Physiological and Acidic Conditions: A Fluorescence Correlation Spectroscopy and Time Resolved Anisotropy Study

  • Suzana M. Andrade
  • Silvia M. B. Costa
  • Jan Willem Borst
  • Arie van Hoek
  • Antonie J. W. G. Visser
Original Paper

Abstract

The interaction between a free-base, anionic water-soluble porphyrin, TSPP, and the drug carrier protein, bovine serum albumin (BSA) has been studied by time-resolved fluorescence anisotropy (TRFA) and fluorescence correlation spectroscopy (FCS) at two different pH-values. Both rotational correlation times and translational diffusion times of the fluorescent species indicate that TSPP binding to albumin induces very little conformational changes in the protein under physiological conditions. By contrast, at low pH, a bi-exponential decay is obtained where a short rotational correlation time (φ int = 1.2 ns) is obtained, which is likely associated to wobbling movement of the porphyrin in the protein binding site. These physical changes are corroborated by circular dichroism (CD) data which show a 37% loss in the protein helicity upon acidification of the medium. In the presence of excess porphyrin formation of porphyrin J-aggregates is induced, which can be detected by time-resolved fluorescence with short characteristic times. This is also reflected in FCS data by an increase in molecular brightness together with a decrease in the number of fluorescent molecules passing through the detection volume of the sample.

Keywords

Porphyrin Albumin Aggregation Binding TRFA FCS 

Notes

Acknowledgements

We thank Dr. J.A.B. Ferreira for helpful discussions on FCS. Professor J. Costa Pessoa is acknowledged for the use of CD spectrometer. This work was supported by POCTI/QUI//57387/2004. S.M. Andrade thanks FCT for the award of Post-Doc grant BPD/24367/2005.

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Copyright information

© Springer Science+Business Media, LLC 2008

Authors and Affiliations

  • Suzana M. Andrade
    • 1
  • Silvia M. B. Costa
    • 1
  • Jan Willem Borst
    • 2
  • Arie van Hoek
    • 2
  • Antonie J. W. G. Visser
    • 2
  1. 1.Centro de Química Estrutural, Complexo 1Instituto Superior TécnicoLisbonPortugal
  2. 2.Wageningen UniversityMicroSpectroscopy CentreWageningenThe Netherlands

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