NMR detection of foreign PAS domain ligands
View Patent ↗Specific binding of a foreign core ligand to a PAS domain, wherein the PAS domain is predetermined, prefolded in its native state, and comprises a hydrophobic core that has no NMR-apparent a priori formed ligand cavity, is determined using NMR spectra of the PAS domain in the presence and absence of a foreign ligand bound within the hydrophobic core. A functional surface binding specificity of a PAS domain is changed by (a) introducing into the hydrophobic core of the PAS domain a foreign ligand of the PAS domain; and (b) detecting a change in the functional surface binding specificity of the PAS domain.
1. A method of detecting binding of a PAS (Per-ARNT-Sim) domain of a protein with a foreign core ligand of the PAS domain, wherein the PAS domain is prefolded in its native state, the method comprising the steps of:
determining from NMR analysis of the PAS domain that the PAS domain comprises a hydrophobic core that has no NMR-apparent a priori formed ligand cavity;
detecting a first NMR spectrum of the PAS domain in the presence of the foreign ligand;
comparing the first NMR spectrum with a second NMR spectrum of the PAS domain in the absence of the ligand; and therefrom
determining the presence of the ligand specifically bound within the hydrophobic core of the PAS domain.
2. The method of claim 1 , wherein the PAS domain is the PAS A domain of PAS kinase.
3. The method of claim 1 wherein the protein consists of the PAS domain.