Lenka, D. R.; Chaurasiya, S.; Kumar, A.

Parkin mutations, including K211N, lead to the onset of Parkinson\'s disease. Ubiquitin-like proteins (NEDD8, Ubiquitin) and Ubiquitin-like (Ubl) domains of Parkin are phosphorylated by PINK1. Parkin is activated by binding of phospho-ubiquitin, phospho-NEDD8, and phospho-Ubl domain on the two distinct pockets of Parkin. While pUb/pNEDD8 binds in the RING1 pocket, pUb/pUbl binding with the RING0 pocket is reported. pUb binding in the RING0 pocket (comprising K211) is believed to rationalize the loss of pUb-mediated Parkin K211N activation. Herein, we demonstrate that the RING0 pocket is specific for pUbl and does not bind with pUb/pNEDD8. In contrast, the RING1 pocket is more suitable for binding with phospho-NEDD8 than phospho-ubiquitin. Also, we demonstrate that the binding of activators in the RING1 and RING0 pockets of Parkin leads to a distinct extent of conformational changes on the RING2. We also show that loss of Parkin activity in disease mutation K211N is independent of Parkin activators (pUb/pNEDD8) binding. Furthermore, the crystal structure of Parkin K211N in complex with pNEDD8 reveals the mechanism of Parkin inactivation due to conformational changes locking RING2 with RING0. This study would help design small-molecule Parkin activators against Parkinson\'s disease.