Abstract
We will discuss several examples from our laboratory of the application of fluorescence correlation spectroscopy (FCS) to studies on protein-ligand interactions in free solution. To demonstrate the usefulness of FCS in such investigations we present situations which span both ligand size (from small molecules of molecular weight MW ≈ lkDa, to large protein ligands) and affinity (picomolar to micromolar).Furthermore, since FCS technology within the biological research community is available in only a small number of labs, we compare the results obtained by FCS with those obtained on the same samples by fluorescence polarization (FP). Such a comparison is useful because FP is a general and well-established optical biophysical tool for homogeneously monitoring molecular associations. While there clearly is overlap in the types of problems amenable to FCS and FP, the information they provide is complementary, and we believe it will be desirable to use them in tandem whenever feasible. We begin by briefly reviewing and comparing FCS and FP.
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Matayoshi, E., Swift, K. (2001). Applications of FCS to Protein-Ligand Interactions: Comparison with Fluorescence Polarization. In: Fluorescence Correlation Spectroscopy. Springer Series in Chemical Physics, vol 65. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-59542-4_5
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DOI: https://doi.org/10.1007/978-3-642-59542-4_5
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