Author:
Schilling Kevin M.,Jorwal Pooja,Ubilla-Rodriguez Natalia C.,Assafa Tufa E.,Gatdula Jean R. P.,Harris David A.,Millhauser Glenn L.
Abstract
AbstractThe C-terminal domain of cellular prion protein (PrPC) contains two N-linked glycosylation sites, the occupancy of which impacts disease pathology. In this study, we demonstrate that glycans at these sites are required to maintain an intramolecular interaction with the N-terminal domain, mediated through a previously identified copper-histidine tether, which suppresses the neurotoxic activity of PrPC. NMR and EPR spectroscopy demonstrate that the glycans refine the structure of the protein’s interdomain interaction. Using whole-cell patch-clamp electrophysiology, we further show that cultured cells expressing PrP molecules with mutated glycosylation sites display large, spontaneous inward currents, a correlate of PrP-induced neurotoxicity. Our findings establish a structural basis for the role of N-linked glycans in maintaining a non-toxic, physiological fold of PrPC.
Publisher
Cold Spring Harbor Laboratory