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bioRxiv · 10.1101/2020.03.20.999326

Development of Nanobodies as first-in-class inhibitors for the NEDP1 deNEDDylating enzyme

Xirodimas, D.·Abidi, N.·Pugniere, M.·Muyldermans, S.·Trauchessec, H.
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Abstract

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.

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Xirodimas, D., Abidi, N., Pugniere, M., Muyldermans, S., Trauchessec, H.. 2020-03-22. Development of Nanobodies as first-in-class inhibitors for the NEDP1 deNEDDylating enzyme. https://doi.org/10.1101/2020.03.20.999326

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