Using a solid-phase ELISA-based binding assay, we determined that ISM binds αv β5 integrin with a K d around 40 μM, in the range of integrin ECM ligand binding affinities (Supplementary Figure S6).
Using this in vitro binding assay, we determined that PARP-1 binds directly to NEIL1.
For the sequence-specific factors (Ctcf and Rxrα), we determined that binding sites were enriched for their known canonical binding motifs.
After analyzing abacavir's binding mode to B*57 01 in the presence and the absence of endogenous peptides, we determined that co-binding peptides play a significant role in the binding mode of drugs in HLA antigen binding cleft.
Additionally, by plotting the apparent enthalpy of binding versus the ionization enthalpy of each buffer and seeing a slope of approximately zero, we determined that upon binding the various DmsAL peptides at pH 8.0, the DmsD::DmsAL complexes do not have a net gain or loss of protons for any of the peptides.
Finally, we determined that the tetramer binding was MHC specific, incapable of binding collagen-specific hybridomas that were restricted by another MHC (for example, DR4-restricted collagen-specific hybridomas, MF = 462 ± 280, n = 4, P < 0.0001; compared with the MF of 11,400 ± 392 of the DR1-restricted CII-specific hybridomas, n = 9).
From the low-loading limit of this quantity, we determined that the binding energy of the sites present on the bud-like aggregates have a value intermediate between that obtained for adsorption on planar graphite and that measured on the grooves of close-ended bundles on nanotubes for the same adsorbate.
Using luciferase reporter gene analysis and site directed mutations of the AP1 or NFAT [21] binding site on the CXCL8 promoter we determined that both AP1 and NFAT were essential for gene activation by PGF2α and that this was independent of NFκB.
We determined that the NFAT binding site is unique to humans; it arose by point mutation along the lineage separating humans from other great apes.
We determined that MutL-ssDNA binding required ATP and was the greatest at ionic strength below 25 mM (KD = 29 nM) while it dramatically decreases above 100 mM (KD>2 µM).
