Abstract
A new methodology of fluorescence decay analysis by iterative reconvolution is presented. It is based on the recent finding that the statistics of single-photon time-correlated data are best described by a compound Poisson law and requires the recording of a sample of at least 20 decays. Application of multivariate statistical methods to the analysis of the recovered decay parameters results in improved accuracy and better estimation of the uncertainties of mono- and multiexponential decays. If it is, of course, not possible to distinguish unambiguously between discrete components and a continuous distribution of lifetimes, it is, however, possible to determine a higher limit of the width of such a distribution should it be present. With our methodology, the presence of a distribution of lifetimes with a width of approximately 20% of its center value inevitably leads to a failure in the deconvolution procedure, a fact of crucial importance in protein conformational studies, for example.
Full text
PDF
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Bajzer Z., Prendergast F. G. A model for multiexponential tryptophan fluorescence intensity decay in proteins. Biophys J. 1993 Dec;65(6):2313–2323. doi: 10.1016/S0006-3495(93)81325-0. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bajzer Z., Sharp J. C., Sedarous S. S., Prendergast F. G. Padé-Laplace method for the analysis of time-resolved fluorescence decay curves. Eur Biophys J. 1990;18(2):101–115. doi: 10.1007/BF00183269. [DOI] [PubMed] [Google Scholar]
- Faller B., Mely Y., Gerard D., Bieth J. G. Heparin-induced conformational change and activation of mucus proteinase inhibitor. Biochemistry. 1992 Sep 8;31(35):8285–8290. doi: 10.1021/bi00150a023. [DOI] [PubMed] [Google Scholar]
- Livesey A. K., Brochon J. C. Analyzing the distribution of decay constants in pulse-fluorimetry using the maximum entropy method. Biophys J. 1987 Nov;52(5):693–706. doi: 10.1016/S0006-3495(87)83264-2. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mely Y., de Rocquigny H., Piémont E., Déméné H., Jullian N., Fournié-Zaluski M. C., Roques B., Gérard D. Influence of the N- and C-terminal chains on the zinc-binding and conformational properties of the central zinc-finger structure of Moloney murine leukaemia virus nucleocapsid protein: a steady-state and time-resolved fluorescence study. Biochim Biophys Acta. 1993 Jan 15;1161(1):6–18. doi: 10.1016/0167-4838(93)90189-x. [DOI] [PubMed] [Google Scholar]
- Mély Y., Piémont E., Sorinas-Jimeno M., de Rocquigny H., Jullian N., Morellet N., Roques B. P., Gérard D. Structural and dynamic characterization of the aromatic amino acids of the human immunodeficiency virus type I nucleocapsid protein zinc fingers and their involvement in heterologous tRNA(Phe) binding: a steady-state and time-resolved fluorescence study. Biophys J. 1993 Oct;65(4):1513–1522. doi: 10.1016/S0006-3495(93)81222-0. [DOI] [PMC free article] [PubMed] [Google Scholar]