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. Author manuscript; available in PMC: 2009 Apr 1.
Published in final edited form as: Protein Expr Purif. 2007 Dec 4;58(2):184–193. doi: 10.1016/j.pep.2007.11.010

Table I. Binding affinity and FRET parameters for interaction between Cy3-HNF4α and Cy5-his-ACBP or Cy5-ACBP.

Cy3 Quenching Cy5 Sensitized emission
Interacting proteins Kd (nM) R2/3 (Å) Kd (nM) R2/3 (Å)
Cy3-HNF4α + Cy5-his-ACBP 111.4 ± 10.2 73.4 ± 6.5 64.4 ± 4.3 74.0 ± 6.7
Cy3-HNF4α + Cy5-ACBP 137.0 ± 11.3 63.8 ± 2.6 28.4 ± 1.6 58.0 ± 5.9
Cy3-βGalactosidase + Cy5-his-ACBP 5700 ± 450 ≫100 5900 ±520 ≫100

All parameters were calculated from FRET curves as presented in Fig. 5 (Methods). Kd, dissociation constant in nM; R2/3, actual distance between Cy3 donor molecules on HNF4α and Cy5 acceptor molecules on ACBP, in angstroms. SD values from n=3.