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Bound ligands were released and separated from the sCD1d molecules by employing a chloroform:methanol:water extraction protocol similar to that previously described by Bligh and Dyer [25].
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After the free ligand is removed by filtration, the PBP-bound ligand is released from the protein by lowering the pH, extracted with organic solvent and analyzed by gas chromatography (GC) for quantification and gas chromatography-mass spectrometry (GC-MS) for identification of the bound ligand.
The heat capacity increments associated with the binding of the protein ligands and contact interaction between the bound ligands were comparable by value but opposite in sign.
Binding sites of proteins without bound ligands were predicted using COACH.
When TNF ligands are released and bind to their cognate receptors, downstream signaling is hypothesized to be affected by the stimulus, synthesis, degradation, and cleavage of pro-caspases.
It has been suggested that nine-amino acid peptide ligands bind specifically to different MHC proteins and that these ligands are released when the MHC proteins break down (for a review, see Hurst [2009]).
After proteolytic cleavage of the membrane-anchored ligand, these ligands are released from the plasma membrane and enable them to bind to death receptors and trigger their activation [ 40].
The preparation of aqueous solutions of monomer lipophilic ligands in equilibrium with protein bound ligand is difficult.
This result indicated that one azido ligand and two PtIV bound hydroxyl groups were released during irradiation.
In this assay, loosely bound histone is released at lower salt concentrations than tightly bound proteins.
On the other hand, once bound a ligand will be released in a richer receptor area when receptors are clustered thereby allowing a greater rebinding probability.
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