Development of NanoLuc-targeting protein degraders and a universal reporter system to benchmark tag-targeted degradation platforms

Grohmann, C; Magtoto, CM; Walker, JR; Chua, NK; Gabrielyan, A; Hall, M; Cobbold, SA; Mieruszynski, S; Brzozowski, M; Simpson, DS; Dong, H; Dorizzi, B; Jacobsen, AV; Morrish, E; Silke, N; Murphy, JM; Heath, JK; Testa, A; Maniaci, C; Ciulli, A; Lessene, G; Silke, J; Feltham, R

Author(s): Grohmann, C; Magtoto, CM; Walker, JR; Chua, NK; Gabrielyan, A; Hall, M; Cobbold, SA; Mieruszynski, S; Brzozowski, M; Simpson, DS; Dong, H; Dorizzi, B; Jacobsen, AV; Morrish, E; Silke, N; Murphy, JM; Heath, JK; Testa, A; Maniaci, C; Ciulli, A; Lessene, G; Silke, J; Feltham, R;
Details: Publication Year 2022-04-19,Volume 13,Issue #1,Page 2073
Journal Title: Nature Communications
Abstract: Modulation of protein abundance using tag-Targeted Protein Degrader (tTPD) systems targeting FKBP12(F36V) (dTAGs) or HaloTag7 (HaloPROTACs) are powerful approaches for preclinical target validation. Interchanging tags and tag-targeting degraders is important to achieve efficient substrate degradation, yet limited degrader/tag pairs are available and side-by-side comparisons have not been performed. To expand the tTPD repertoire we developed catalytic NanoLuc-targeting PROTACs (NanoTACs) to hijack the CRL4(CRBN) complex and degrade NanoLuc tagged substrates, enabling rapid luminescence-based degradation screening. To benchmark NanoTACs against existing tTPD systems we use an interchangeable reporter system to comparatively test optimal degrader/tag pairs. Overall, we find the dTAG system exhibits superior degradation. To align tag-induced degradation with physiology we demonstrate that NanoTACs limit MLKL-driven necroptosis. In this work we extend the tTPD platform to include NanoTACs adding flexibility to tTPD studies, and benchmark each tTPD system to highlight the importance of comparing each system against each substrate.
Publisher: NPG
Keywords: Benchmarking; Luciferases; Proteolysis; Tacrolimus Binding Protein 1A/genetics
Research Division(s): Chemical Biology; Inflammation; Epigenetics And Development; Ubiquitin Signalling
PubMed ID: 35440107
Publisher's Version: https://doi.org/10.1038/s41467-022-29670-1
Open Access at Publisher's Site: https://doi.org/ 10.1038/s41467-022-29670-1
Terms of Use/Rights Notice: Refer to copyright notice on published article.
Documents: s41467-022-29670-1.pdf - (2.49MB, application/pdf)

Creation Date: 2022-05-03 09:18:31

Last Modified: 2022-05-03 09:27:36