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The crystal structure of human SULT1E1 with PAP and E2 bound to the active site (see Supplemental Material, Figures S1A,B and S2A) is similar in overall fold and substrate binding as previously determined for the mouse estrogen sulfotransferase (Kakuta et al. 1997).
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We have previously determined PP1 as a binding partner with this approach.
As such, should ligand binding deform the protein conformation beyond the previously determined conformation, the control restraints will report the discrepancy by increasing the system's energy.
These very low concentrations were unexpected as a Kd of 2.1 µM had been previously determined for the binding of E2 to rat PDI [5].
Catalytic and substrate-binding residues of sxtA from cyanobacteria have been previously determined [6], [41].
Equilibrium Kd values for the binding of the TnH 30−61) and TnH(126−159) peptides were determined using competition with melittin binding as previously described (26).
Samples were scored to determine the proportion of tumor cells exhibiting pimonidazole binding, as previously described [ 35].
OpuAC from Bacillus subtilis has previously been investigated in terms of ligand binding as well as having its structure determined [15], [16].
The number of receptors and binding affinities were determined as previously described (Obiri et al, 1993; Puri et al, 1991).
In order to determine the binding affinity of the secreted scFvs with the target androgen-independent prostate cancer cells, saturation binding assays were performed essentially as previously described [ 36, 40].
A minimum of two PBM experiments were performed with each purified protein construct to determine their binding specificities as previously described [ 13, 14, 26].
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