Niko Klink

Weta workshop?

Very happy to see our contribution to a better understanding of Hsp70/90 #chaperones online! Congrats and a big thank you to all co-authors, collaborators and reviewers! #proteostasis @poepsel-lab.bsky.social @crc1430.bsky.social @unidue-zmb.bsky.social

The degrader is much much worse in cellular permeability than the inhibitor (we tested), but is just so potent at forming a stable ternary complex and inducing degradation that we just need less for full degradation!

Thanks alot, much appreciated, and thanks for the feedback! Would you like to see inhibitor go down or degrader go up with the concentration? Equimolarity is a bit iffy to compare here, since they have such drastically different membrane permeability

And wishing you a full and speedy recovery!

Ah now I understand your question correctly, i thought you meant something different! Sorry to hear that. I dont know enough about long term treatment of Kras inhibitors to answer that question truthfully, sorry!

Yes, it will very much hinder degradation! The inhibitor has higher cell membrane permeability and will outcompete the Degrader binding site

Thank you! By becoming available later you mean Treating cells with inhibitor first then adding degrader at later time points?

Lastly, a huge thank you to my supervisor Dr. @maltegersch.bsky.social for his guidance and support, as well as the entire Gersch team.
I am very glad to share this story, but even more excited for whats to come in the (near) future, so stay tuned, we are just getting started!
#TPD #PROTAC #DUB

Big thanks also to Dr. Bikash Adhikari and Prof. Elmar Wolf from @wolflab23.bsky.social, as well as Dr. @martinschwalm.bsky.social for bringing their expertise to this project, @imprs-lm.bsky.social from the MPI Dortmund for the excellent support and everyone else who contributed to the prokect!

This work would not have been possible without my amazing co-first authors Dr. Sebastian Urban and Dr. Med. Johanna Seier and everyone in this highly interdisciplinary team from the @crc1430.bsky.social and beyond.

Taking these now to our Model Systems (PDAC and melanoma), not only shows highly selective degradation of USP7 by our PROTACs (criteria for matched pair fulfilled!), but also striking differences between inhibitor and degrader treated cells. These differences also occur in many phenotypic assays!

After making a bunch of degraders (most not shown here), we arrive at NK250 and NK266 ("Kurt Cubane"), two highly potent USP7 degraders in HiBiT and immune-based assays. NK250, is a faster degrader which we could correlate to its ability to form a stable ternary complex between USP7 and VHL.

We introduce NK192, a potent USP7 inhibitor of the widely used hydroxypiperidine scaffold. Converting it through the available exit vector to a biotin probe showed proteome-wide highly selective binding to USP7, fulfilling our criteria for the inhibitor side of the matched chemical pairs.

To enable their systematic discovery, we addressed this urgent need with matched chemical pairs of DUB inhibitor and #degraders by focusing on USP7 as a case study. By using potent and selective modulators, we aimed to dissect phenotypes induced by inhibitors and degraders of USP7 in solid cancers.

#Deubiquitinases (DUBs), key regulators of ubiquitin signaling, have been increasingly recognized for functions beyond rescuing substrate proteins from proteasomal degradation. Due to a lack of selective chemical tools within the DUB space, finding these functions has proven to be very challenging.

Targeted degradation of USP7 in solid cancer cells reveals disparate effects of deubiquitinase inhibition vs. acute protein depletion Proteolysis-targeting chimeras (PROTACs) co-op the ubiquitin system for targeted protein degradation, creating opportunities to interrogate cellular functions of proteins through "chemical knockdown"....

Really excited to share the first chapter of my PhD as a preprint titled: Targeted degradation of USP7 in solid cancer cells reveals disparate effects of deubiquitinase inhibition vs. acute protein depletion: www.biorxiv.org/content/10.1.... If that sounds interesting to you, small tweetorial below👇

We hope that our work paves the way for the next generation of molecules targeting USP30 and fighting neurodegenerative diseases, and hope that this strategy will soon be applied to other DUBs!

Huge shoutout to Nafizul Kazi for this incredible tour de force on tireless protein engineering and thorough biochemical assay characterizations, @gallantkai.bsky.social and Gian Marvin Kipka for the beautiful cellular work and @maltegersch.bsky.social for amazing guidance and supervision.

Chimeric deubiquitinase engineering reveals structural basis for specific inhibition of the mitophagy regulator USP30 - Nature Structural & Molecular Biology Kazi et al. report the crystal structure of the mitochondrial deubiquitinase USP, a clinical stage Parkinson’s disease drug target, in complex with a specific inhibitor. The authors delineate a framew...

Very excited to share that our work on Parkinson's target USP30 has been published in Nature Structural & Molecular Biology! By combining soluble DUB-domains onto USP30, we for the first time achieved co-crystallization with a small molecule (NK036) without Nanobodies! www.nature.com/articles/s41...

Absolutely fantastic work, congrats to the whole team!

In-cell architecture of the mitochondrial respiratory chain Mitochondria regenerate adenosine triphosphate (ATP) through oxidative phosphorylation. This process is carried out by five membrane-bound complexes collectively known as the respiratory chain, workin...

Wow. This is really stunning . www.science.org/doi/10.1126/...

Ubiquitin is directly linked via an ester to protein-conjugated mono-ADP-ribose | The EMBO Journal imageimageCertain E3 ligases have been found to ubiquitylate hydroxyl groups on free NAD+ and ADP-ribose in vitro, but the in vivo occurrence of this dual post-translational modification has remained ...

I'm super excited to share our study in collaboration with @jnpruneda.bsky.social, published today in @embojournal.org! We show that PARPs are modified with a non-canonical ester-linked mono-ADP-ribosylated ubiquitin species for the first time in cells: www.embopress.org/doi/full/10....

C-terminal amides mark proteins for degradation via SCF–FBXO31 - Nature SCF–FBXO31 scans proteins for C-terminal amidation and marks them for subsequent proteasomal degradation.

🎉Super excited to share our story on how the substrate receptor FBXO31 functions as a quality control factor by recognizing amides. This has been an amazing collaboration between Bode lab and @jcornlab.bsky.social. Special shutout goes to @matthiasmuhar.bsky.social www.nature.com/articles/s41...

I dont recall where I read it, but dont These have rather Poor affinity for CRBN compared to the usual suspects?

Congrats Georg, fantastic news and well deserved!

A fantastic story from our lab about finding novel activity in two "inactive" DUBs!

go.bsky.app/3VPQofn

Great way to get started here!