RT Journal Article
SR Electronic
T1 Development of Nanobodies as first-in-class inhibitors for the NEDP1 deNEDDylating enzyme
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 2020.03.20.999326
DO 10.1101/2020.03.20.999326
A1 Naima Abidi
A1 Helene Trauchessec
A1 Gholamreza Hassanzadeh-Ghassabeh
A1 Martine Pugniere
A1 Serge Muyldermans
A1 Dimitris P. Xirodimas
YR 2020
UL http://biorxiv.org/content/early/2020/03/23/2020.03.20.999326.abstract
AB Protein NEDDylation emerges as an important post-translational modification and an attractive target for therapeutic intervention. Modification of NEDD8 onto substrates is finely balanced by the co-ordinated activity of conjugating and deconjugating enzymes. The NEDP1/DEN1/SENP8 protease is a NEDD8 specific processing and deconjugating enzyme that regulates the NEDDylation mainly of non-cullin substrates. Here, we report the development and characterisation of nanobodies as first-in-class inhibitors for NEDP1. The nanobodies display high-affinity (low nM) against NEDP1 and specifically inhibit NEDP1 processing activity in vitro and NEDP1 deconjugating activity in tissue-culture cells and in cell extracts. We also isolated nanobodies that bind to NEDP1 with high-affinity but do not affect NEDP1 activity. The developed nanobodies provide new tools to study the function of NEDP1 and to prevent deNEDDylation in cell extracts used in biochemical assays.