Fig. 3: Surface plasmon resonance (SPR) and isothermal titration calorimetry analysis of SHP-2 interaction with PD-1.
From: Interaction of SHP-2 SH2 domains with PD-1 ITSM induces PD-1 dimerization and SHP-2 activation
a PD-1 phosphotyrosyl ITSM-Y248 peptide was immobilized on a CM5 BIAcore chip and binding of SHP-2 full-length SHP-2-N-SH2, and SHP-2-C-SH2, all at 320 nM, was determined by SPR. Full binding curves are presented in Supplementary Fig.  8. Sensograms of 900 s (15 min) association and 900 s dissociation time were analyzed and KD values were calculated. b PD-1 phosphotyrosyl ITIM-Y223 peptide was immobilized on a CM5 BIAcore chip and binding of SHP-2-full-length at the indicated concentrations, was determined by SPR. Results are representative of three experiments. c, d Isothermal titration calorimetry (ITC) experiments were performed on a MicroCal ITC200 instrument. PD-1 phosphopeptides (PD-1cyto-pITIM-pITSM and PD-1cyto-ITIM-pITSM) and t-SHP-2 protein were prepared in buffer containing 50 mM HEPES pH 7.4, 100 mM NaCl, 5 mM TCEP and stoichiometry of bisphosphorylated PD-1 peptide: t-SHP-2 interaction was assessed as described in Methods.
