Inferring Diffusion Dynamics from FCS in Heterogeneous Nuclear Environments

Konstantinos Tsekouras, Amanda P. Siegel, Richard Day, Steve Pressé

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Abstract Fluorescence correlation spectroscopy (FCS) is a noninvasive technique that probes the diffusion dynamics of proteins down to single-molecule sensitivity in living cells. Critical mechanistic insight is often drawn from FCS experiments by fitting the resulting time-intensity correlation function, G(t), to known diffusion models. When simple models fail, the complex diffusion dynamics of proteins within heterogeneous cellular environments can be fit to anomalous diffusion models with adjustable anomalous exponents. Here, we take a different approach. We use the maximum entropy method to show - first using synthetic data - that a model for proteins diffusing while stochastically binding/unbinding to various affinity sites in living cells gives rise to a G(t) that could otherwise be equally well fit using anomalous diffusion models. We explain the mechanistic insight derived from our method. In particular, using real FCS data, we describe how the effects of cell crowding and binding to affinity sites manifest themselves in the behavior of G(t). Our focus is on the diffusive behavior of an engineered protein in 1) the heterochromatin region of the cell's nucleus as well as 2) in the cell's cytoplasm and 3) in solution. The protein consists of the basic region-leucine zipper (BZip) domain of the CCAAT/enhancer-binding protein (C/EBP) fused to fluorescent proteins.

Original languageEnglish
Article number6597
Pages (from-to)7-17
Number of pages11
JournalBiophysical Journal
Volume109
Issue number1
DOIs
StatePublished - Jul 9 2015

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Fluorescence Spectrometry
Proteins
CCAAT-Enhancer-Binding Proteins
Leucine Zippers
Entropy
Cell Nucleus
Cytoplasm

ASJC Scopus subject areas

  • Biophysics

Cite this

Inferring Diffusion Dynamics from FCS in Heterogeneous Nuclear Environments. / Tsekouras, Konstantinos; Siegel, Amanda P.; Day, Richard; Pressé, Steve.

In: Biophysical Journal, Vol. 109, No. 1, 6597, 09.07.2015, p. 7-17.

Research output: Contribution to journalArticle

Tsekouras, Konstantinos ; Siegel, Amanda P. ; Day, Richard ; Pressé, Steve. / Inferring Diffusion Dynamics from FCS in Heterogeneous Nuclear Environments. In: Biophysical Journal. 2015 ; Vol. 109, No. 1. pp. 7-17.
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