Anomalous Negative Fluorescence Anisotropy in Yellow Fluorescent Protein (YFP 10C): Quantitative Analysis of FRET in YFP Dimers

. Author manuscript; available in PMC: 2008 Dec 18.

Published in final edited form as: Biochemistry. 2007 Nov 21;46(50):14403–14417. doi: 10.1021/bi701575n

In order to obtain the YFP dissociation constant K_d from the concentration dependence of excitation spectra (Fig. 2C), we analyzed the contributions of the various components of our system, which involves two oligomerization states of the protein and two protonation states of the chromophore. Combinations of these two properties result in the multiple species present in YFP at pH 6.0 as listed below

Name	Definition
mYFP_A	monomeric YFP with a neutral chromophore
mYFP_B	monomeric YFP with an anionic chromophore
dYFP_AA	dimeric YFP with two neutral chromophores
dYFP_BB	dimeric YFP with two anionic chromophores
dYFP_AB	dimeric YFP with a neutral and an anionic chromophore