Dynamics of ligand binding from 13C NMR relaxation dispersion at natural abundance.
We show that Carr-Purcell-Meiboom-Gill (CPMG) 13Calpha NMR relaxation dispersion measurements are a viable means for profiling mus-ms ligand dynamics involved in receptor binding. Critically, the dispersion is at natural 13C abundance; this matches typical pharmaceutical research settings in which ligand isotope-labeling is often impractical. The dispersion reveals ligand 13Calpha nuclei that experience mus-ms modulation of their chemical shifts due to binding. 13Calpha shifts are dominated by local torsion angles , psi, chi1; hence, these experiments identify flexible torsion angles that may assist complex formation. Since the experiments detect the ligand, they are viable even in the absence of a receptor structure. The mus-ms dynamic information gained helps establish flexibility-activity relationships. We apply these experiments to study the binding of a phospho-peptide substrate ligand to the peptidyl-prolyl isomerase Pin1.
Department of Chemistry and Biochemistry, University of Notre Dame, 251 Nieuwland Science Hall, Notre Dame, Indiana 46556, USA.
John S
JS
Zintsmaster
Brian D
BD
Wilson
Jeffrey W
JW
Peng
D001665:Binding Sites; D002247:Carbon Isotopes; D006801:Humans; D008024:Ligands; D009682:Magnetic Resonance Spectroscopy / Q000379:methods*; D008956:Models, Chemical; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D019696:Peptidylprolyl Isomerase / Q000737:chemistry*; D010748:Phosphopeptides / Q000737:chemistry*
D002247:Carbon Isotopes; D008024:Ligands; D000072340:NIMA-Interacting Peptidylprolyl Isomerase; D010748:Phosphopeptides; C000606846:PIN1 protein, human; D019696:Peptidylprolyl Isomerase