Chemical biology
PROTAC THNAN69 degrades LIMK2, not LIMK1
Open access · cc by · source: Europe PMC
A cereblon-recruiting degrader based on LIMKi3 and a phenyl dihydrouracil ligand wipes out LIMK2 at nanomolar DC50 while leaving LIMK1 and phospho-cofilin largely intact.
Study at a glance
- Design
- Animal / in-vitro — CRBN/VHL PROTACs of LIMKi3 ranked by cellular degradation and proteomics
- N
- Cell-based degrader SAR (e.g., HuCCT1 EGFP–LIMK2 / Western blot) — no subject N
- Population
- Cultured cells expressing LIMK2 (including HuCCT1)
- Outcome
- LIMK2-selective degradation potency (THNAN69 DC50 ≈ 1 nM)
Structured fields used in claim comparison tables when every cited study has a complete layer.
Key findings
THNAN69 DC50 = 1 nM (vs 33 nM for thalidomide analogue 21b); 10 nM depletes endogenous LIMK2. Proteome-wide, LIMK2 was the only kinase degraded. VHL PROTACs bound poorly in cells. Cofilin phosphorylation persisted via LIMK1 compensation.
Methodology
Authors made CRBN and VHL PROTACs of LIMKi3, ranked them by DSF, SPR, NanoBRET, EGFP–LIMK2 live degradation, Western blot, and proteomics, then used MD of ternary complexes to explain isoform bias.
Limitations
No animal efficacy or clinical data; phospho-cofilin does not fall, so LIMK2-only degradation is not a cofilin-pathway knockout.
How this study connects
Role on claims
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This library holds 8 empirical chemistry papers on chemical biology with isolated findings, rates or spectra rather than reviews.
Evidence for the claim as stated.
A cereblon-recruiting degrader based on LIMKi3 and a phenyl dihydrouracil ligand wipes out LIMK2 at nanomolar DC50 while leaving LIMK1 and phospho-cofilin largely intact.
Evidence for the claim as stated.
A binding pose or a DC50 in one cell line is not a drug; off-target proteome labelling varies with the tag.
Evidence for the claim as stated.
HPLC-adjacent assays in this set also include enzyme activity workups. E. coli DERA is irreversibly inactivated after 200–300 mM acetaldehyde; crotonaldehyde inhibits more than 100-fold more potently (1 mM, <1 h) via K167/C47; C47M keeps 60 ± 5% activity and loses none after 16 h in 300 mM acetaldehyde. LIMK2 PROTAC THNAN69 has DC₅₀ = 1 nM (versus 33 nM for analogue 21b) and 10 nM depletes endogenous LIMK2 — proteomics/Western, not a simple UV chromatogram, carry that selectivity claim.
Evidence for the claim as stated.
Open questions
Tensions this paper is part of
From concept pages' “where studies disagree.” Disagreement means the same question; scope means different assays, populations, or outcomes.
A binding pose or a DC50 in one cell line is not a drug; off-target proteome labelling varies with the tag.
Related papers in this topic
Same topic cluster — not a recommendation engine.