Protein kinases are critical regulators of cellular signaling, but precise modulation of their activity remains challenging due to their high structural conservation. Here, we present de novo designed genetically encoded miniproteins capable of activating or inhibiting focal adhesion kinase (FAK) by directly targeting the kinase domain itself. Among 96 binders designed to stabilize distinct conformational states of FAK, 33 modulated kinase activity. Biochemical characterization of the four most potent modulators revealed that two designs inhibit FAK with low-nanomolar IC50 values while the remaining two potentiated FAK activity by more than two-fold. When expressed in cells, the modulators preserved the same inhibitory and activating effects observed in vitro, establishing that designed conformational binders can directly tune FAK signaling in living cells. Taking advantage of the high similarity between kinases, we redesigned the FAK inhibitors to inhibit Src kinase. Our approach establishes a versatile platform for selective and genetically encoded kinase control as a way to rewire cell signaling and as a starting point for the discovery of novel modulatory sites of kinases.
Together, these examples define an expanding pharmacological vocabulary for kinase drug discovery: one that asks not only whether a molecule inhibits its target kinase, but how ligand binding rewires protein stability, interactions, localisation, and function.
J. Diepeveen, Z. Kozicka· Chimia (Basel)· 0 citations
The structural and medicinal chemistry principles underlying (i) allosteric inhibition and (ii) proximity-induced degradation are summarized, with an emphasis on design logic, structure-activity relationships, and key liabilities in the beyond rule of five space.
Mei Zhou, Linshan Li, Xiao-Juan Tang et al.· Future Medicinal Chemistry· 0 citations
Protein phosphorylation is dynamically controlled by kinases and phosphatases, and its dysregulation contributes to diverse disease-relevant states. Although conventional kinase modulators have enabled important therapeutic advances, direct kinase or phosphatase modulation often lacks the precision needed to correct phosphorylation at the level of a defined protein of interest (POI). Proximity-inducing modalities, particularly phosphorylation-inducing chimeric small molecules (PHICSs) and phosphatase-recruiting chimeras (PHORCs), offer an event-driven strategy to modulate phosphorylation by recruiting catalytic effectors to selected targets. Recent studies have extended PHICSs beyond early proof-of-concept systems, highlighting both improved pan-AMPK recruitment strategies and self-recruiting designs that redirect oncogenic kinase activity toward inhibitory phosphorylation. In parallel, recent PHORC studies have diversified induced dephosphorylation, spanning tag-based signaling rewiring, simultaneous PP5 recruitment and activation, and aptamer-guided PTPRF recruitment for receptor regulation. Together, these studies highlight the expanding scope of proximity-induced phosphorylation control, while emphasizing that broader application will depend on improved molecular design and a clearer understanding of proximity-driven mechanisms.
Yuxin Xia, Daichao Zhai, Qi-Dong You et al.· RSC Medicinal Chemistry· 0 citations
Members of the DYRK kinase family have been implicated in the regulation of key metabolic processes. In particular, DYRK1B plays a central role in hepatic lipid metabolism and fatty liver disease. Notably, these functions depend not only on the enzyme’s catalytic activity but also on scaffolding properties. Consequently, therapeutic ambitions aiming to target the entire spectrum of DYRK1B functions must inactivate or eliminate the protein itself rather than merely inhibiting its enzymatic activity, as is achieved by current kinase inhibitors. In this study, we describe a small molecule from the leucettinib class of DYRK inhibitors (LCTB-92) that not only suppresses DYRK kinase activity but also promotes selective ablation of the DYRK1B protein while sparing the closely related paralog DYRK1A. The identification of this compound establishes a foundation for the development of selective agents capable of targeting the entire DYRK1B protein scaffold, thereby addressing catalytic-dependent as well as catalytic-independent kinase functions.
Miriam S. Ems, Mattias F. Lindberg, Till Adhikary et al.· ACS Pharmacology & Trans...· 0 citations
The activity and abundance of >160,000 variants are quantify the activity and abundance of >160,000 variants to construct complete maps of the energetic and allosteric architectures of five human kinase domains: SRC, FGR, JNK2/MAPK9, ZAK/MAP3K20, and TSSK2.
Carla Folgado, Antoni Beltran, Ben Lehner· bioRxiv· 0 citations
A fundamental question in cell signaling is how a single kinase, with a single primary substrate, can activate multiple distinct cellular programs. Kinases can be characterized by speed, duration, and amplitude. They should be efficient—but not necessarily maximally—as signaling functions often require slow kinetics. How the physical behavior of individual kinases, especially the top-tier kinase, coordinates the collective cascade for functional output—essential for molecular engineering and pharmacology—remains a challenge. Here we decipher the structural “why” behind the specific kinase order in cascades as the key design principle of cellular signaling, which resolves these questions. We elucidate the conformational mechanisms and catalytic actions of the sequentially ordered component kinases in seven related and distinct cascades, including MAPKs. We propose that the order of kinases in cascades is dictated by their conformational character, a mechanism amplified in cascade-enriched biomolecular condensates—establishing design principles for pathway-selective, cascade-directed therapeutics.
R. Nussinov, Clil Regev, H. Jang· RSC Chemical Biology· 0 citations
We use cookies to run the site and, with your consent, for analytics and to show ads.
See our Cookie Policy.
