Structures of other complexes of 14-3-3 14-3-3 14-3-3roteins have been determined as with,37,38,39,40 but the binding sites for these proteins show less overlap with the ExoS/T-binding site than that for trehalase (Supplementary Fig. 15).
A precise region of 79 amino acids, representing less than 2.5% of the entire protein, was determined as its binding site.
Binding was determined as 1 1 through identical binding capacity of immobilized RGL1N for recombinant GID1A/GA4 and the monoclonal antibody BC9 (results not shown).
Specific [3H]-DEX binding was determined as the difference between total and nonspecific binding and was expressed as sites/cell.
A. castellanii (106 amoebae/well) were pre-incubated with various concentrations of Sn(IV porphyrin or solvent alone for 45 min under visible light and then added to cell monolayers for 1 h and percentage binding determined as follows: 100 – [no. of unbound amoebae/total number of amoebae x 100] =% bound amoebae.
The percentage FAM-labeled St-2-1 replaced by the tested aptamer was determined as a measure of binding affinity for the aptamer (Table 1).
Using the same method as described above to obtain the mean values and the standard errors of the mean from three repeated experiments for Δ H°, Ka, n, − TΔ S°, and Δ G° for CaM Fas DD WT binding, the thermodynamic parameters for CaM Fas DD V254N binding were determined as shown in Table 1.
Subsequently a one-site binding model was determined as the appropriate form of analysis for all binding data.
Total binding corresponds to the toxin bound after 100 min of association, and the irreversible binding was determined as the bound toxin that remains after 60 min under dissociation conditions.
Within the context of a CWR22Pc-R1-AD1 CWR22Pc-R1-AD1 CWR22Pc-R1-AD1ively expresses a strucellally full-length AR containing a H874Y mutation23, the rank order of antagonist backgrounds determined as ARN-509 > Hydroxyflutamide > Enzalutamide > Nilutamide > Bicalutamide (Fig. 3c).
