Extended Data Fig. 6: NMR analysis of ALK-ECRABR–ALKAL2-AD and ALK-ECRABR–ALKAL1-AD complexes.
From: Mechanism for the activation of the anaplastic lymphoma kinase receptor
a, b, Superimposed 1H-13C–correlated spectra of ALK-ECRABR–ALKAL2-AD (a) and ALK-ECRABR–ALKAL1-AD (b) complexes. ALK-ECRABR and ALKAL proteins are 1H-13C labelled in the indicated methyl groups. c, d, Chemical shift perturbation induced by ALKAL1-AD binding to ALK-ECRABR to combined 1H and 15N amide atoms (c) and 1H and 13C methyl atoms (d). e, f, Chemical shift perturbation induced by ALKAL1-AD mapped onto the ALK-ECRABR structure. g, NMR characterization of ALKAL1-AD binding to ALK-ECRABR. Select strips from 13C-edited NOESY experiments showing intermolecular NOEs between ALK-ECRABR and ALKAL1-AD. Similar results were obtained when ALKAL2-AD was used, confirming that the structure observed in the frozen sample used in cryo-EM is the same in solution. h, NMR characterization of the EGF-like domain repositioning upon ALKAL1 binding to ECRABR. Select strips from 13C-edited NOESY experiments for ALK-ECRABR showing interdomain NOEs in the unbound form. Characteristic NOE patterns between Met997 of the EGF-like domain and the indicated residues of the TNF-like domain (right panel) changed dramatically upon ligand binding and demonstrate pronounced re-orientation of the EGF-like domain as shown schematically on the right panel.
