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Covalent Reprogramming of Kinase Binders to Modulate Protein Homeostasis

ABSTRACT: Small molecules that modulate protein abundance through induced proximity have expanded the landscape beyond traditional inhibition. Here, we explore how introducing covalent or latent electrophilic groups into a multi-kinase binder scaffold can reprogram protein homeostasis within the kinase family. Using the broad-spectrum kinase ligand TL13-87 as a template, we synthesized analogs bearing α-chloroacetamide, acrylamide, or terminal amine groups. Quantitative proteomics revealed that while most analogs had minimal global impact, MKI-AA uniquely stabilized the mitotic kinase AURKA, a protein often destabilized by ATP-competitive inhibitors. Mechanistic studies showed that MKI-AA acts post-translationally to suppress AURKA ubiquitination and proteasomal degradation. Proteomic mapping of MKI-AA-induced AURKA interactors revealed changes in protein associations upon treatment, providing potential mechanistic insights into how MKI-AA influences AURKA stability. Intriguingly, adding a short linker to MKI-AA converted it from a stabilizer into a degrader, highlighting how subtle structural variations can invert functional outcomes. These findings demonstrate that electrophilic ligand design can modulate kinase stability and reveal a previously unrecognized mode of covalent proximity-driven protein stabilization.

INSTRUMENT(S):

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Xiaoyu Zhang

LAB HEAD: Xiaoyu Zhang

PROVIDER: PXD071390 | Pride | 2026-04-02

REPOSITORIES: Pride

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			Action	DRS
	293T_MKDs_WP_MS3_10plex_01.raw	Raw
	293T_MKDs_WP_MS3_10plex_02.raw	Raw
	293T_MKDs_WP_MS3_10plex_03.raw	Raw
	293T_MKDs_WP_MS3_10plex_04.raw	Raw
	293T_MKDs_WP_MS3_10plex_05.raw	Raw

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