Identification of actionable targeted protein degradation effector sites through Site-specific Ligand Incorporation-induced Proximity (SLIP)

Опубликована: Фев. 4, 2025

ABSTRACT Targeted protein degradation (TPD) is a rapidly emerging and potentially transformative therapeutic modality. However, the large majority of >600 known ubiquitin ligases have yet to be exploited as TPD effectors by proteolysis-targeting chimeras (PROTACs) or molecular glue degraders (MGDs). We report here chemical–genetic platform, Site-specific Ligand Incorporation-induced Proximity (SLIP), identify actionable (“PROTACable”) sites on any potential effector in intact cells. SLIP uses genetic code expansion (GCE) encode copper-free “click” ligation at specific site cells, enabling situ formation covalent PROTAC-effector conjugate against target interest (POI). Modification drives targeted protein, establishing these for TPD. Using SLIP, we systematically screened dozens across E3 E2 enzymes from diverse classes, identifying multiple novel PROTACable which are competent adds powerful approach proximity-induced pharmacology (PIP) toolbox, future ligand discovery fully enable TPD, other PIP modalities.

Язык: Английский

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E3 ubiquitin ligases and their therapeutic potential in disease Management

Biochemical Pharmacology, Год журнала: 2025, Номер unknown, С. 116875 - 116875

Опубликована: Март 1, 2025

Язык: Английский

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Multilayered HIV-1 resistance in HSPCs through CCR5 Knockout and B cell secretion of HIV-inhibiting antibodies

Nature Communications, Год журнала: 2025, Номер 16(1)

Опубликована: Апрель 1, 2025

Allogeneic transplantation of CCR5 null hematopoietic stem and progenitor cells (HSPCs) is the only known cure for HIV-1 infection. However, this treatment limited because rarity CCR5-null matched donors, morbidities associated with allogeneic transplantation, prevalence strains resistant to knockout (KO) alone. Here, we propose a one-time therapy through autologous HSPCs genetically engineered ex vivo produce both KO long-term secretion potent inhibiting antibodies from B cell progeny. CRISPR-Cas9-engineered engraft reconstitute multiple lineages in can be express simultaneously (in pre-clinical models). Human each antibody secrete neutralizing concentrations capable pseudovirus infection vitro. This work lays foundation potential functional combining delivery therapeutic against efficacy HSPC transplantation. Transplantation (HSCs) reported HIV-1. authors describe an HSC transplant multilayered resistance HIV by

Язык: Английский

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Targeted protein degradation for cancer therapy

Nature reviews. Cancer, Год журнала: 2025, Номер unknown

Опубликована: Апрель 25, 2025

Язык: Английский

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Illuminating ubiquitination mechanisms: How cryo-EM has shed light on Cullin RING E3 ligase function

Current Opinion in Structural Biology, Год журнала: 2025, Номер 92, С. 103055 - 103055

Опубликована: Май 1, 2025

The ubiquitin-proteasome system (UPS) governs protein homeostasis by orchestrating the selective degradation of regulatory and misfolded proteins through a tightly regulated series ATP-driven ubiquitination reactions. E3 ubiquitin ligases play central role in this process conferring substrate specificity, yet structural complexity dynamic nature these large macromolecular assemblies poses challenges for traditional biology techniques such as X-ray crystallography nuclear magnetic resonance (NMR). advent single-particle cryo-electron microscopy (cryo-EM) has transformed our ability to study enzymes, revealing previously inaccessible mechanistic insights into their allosteric regulation, conformational transitions, recognition. By integrating high-resolution crystallographic data with cryo-EM's resolve heterogeneous complexes, researchers have uncovered fundamental principles governing ligase activity. This review explores how cryo-EM reshaped understanding Ligases. We highlight key discoveries enabled technique, discuss emerging approaches, alongside complementary methodologies, are advancing therapeutic strategies targeting signaling family ligases.

Язык: Английский

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Drug discovery technologies–Current and future trends

Elsevier eBooks, Год журнала: 2025, Номер unknown

Опубликована: Янв. 1, 2025

Язык: Английский

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Primed for degradation: How weak protein interactions enable molecular glue degraders

Current Opinion in Structural Biology, Год журнала: 2025, Номер 92, С. 103052 - 103052

Опубликована: Май 6, 2025

Molecular glues are small drug-like molecules that induce de novo protein-protein interactions or facilitate pre-existing weak between proteins. In the context of a ubiquitin ligase, such binding events frequently result in ubiquitination by proximity. Rational development these transformative modalities, however, remains major challenge. Here we review recent insights into molecular and emerging design principles. Protein surfaces can similarly be complemented mutations compounds inducing resulting gain functionality. When interaction surface two proteins is relatively small, when affinity otherwise weak, proportionally more energy will have to provided compound glue together. We suggest simple thermodynamic model rationalize action facilitated mutations.

Язык: Английский

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Degradation Bottlenecks and Resource Competition in Transiently and Stably Engineered Mammalian Cells

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июнь 3, 2024

Abstract Degradation tags, otherwise known as degrons, are portable sequences that can be used to alter protein stability. Here, we report degron-tagged proteins compete for cellular degradation resources in engineered mammalian cells leading coupling of the rates independently expressed when constitutively targeted human degrons adopted. By adopting inducible bacterial and plant also highlight how orthogonality uncoupling synthetic construct from native machinery achieved. We show effect this competition dependent on context where C-terminal appears impact most across our tested settings. then build a genomically integrated capacity monitor tagged with different confirm resource between genomic transiently DNA constructs. This work expands characterisation including systems, providing framework optimisation heterologous expression systems advance applications fundamental applied biological research.

Язык: Английский

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Lessons from natural molecular glue degraders

Biochemical Society Transactions, Год журнала: 2024, Номер 52(3), С. 1191 - 1197

Опубликована: Июнь 12, 2024

Molecular glue (MG) degraders include plant hormones and therapeutic drugs have become a hot topic in drug discovery. Unlike bivalent proteolysis targeting chimeras (PROTACs), monovalent MGs can trigger the degradation of non-ligandable proteins by enhancing their interaction with E3 ubiquitin ligases. Here, I analyze characteristics natural MG degraders, contrast them synthetic ones, provide rationale for optimizing MGs. In MG-based systems, stable complex is only formed when all three partners (MG, ligase, substrate) are present, while affinities between any two components either weak or undetectable. After substrate degraded, will dissociate from its receptor (E3 ligase) due to low micromolar affinity. contrast, MGs, such as immunomodulatory (IMiDs) CR8, potent inhibitors receptors blocking CRBN-native occupying active site CDK12. Inspired nature, IMiDs CRBN be reduced make those compounds without E3-inhibitory activity, therefore, minimizing interference physiological substrates CRBN. Similarly, CR8-CDK weakened uncouple degrader function kinase inhibition. To mimic examples reduce side effects, future development that lack inhibitory activity should considered.

Язык: Английский

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Crispr-based gene therapy for the induction of fetal hemoglobin in sickle cell disease

Expert Review of Hematology, Год журнала: 2024, Номер unknown

Опубликована: Ноя. 13, 2024

Sickle cell disease is ameliorated, and perhaps can be 'cured' if enough fetal hemoglobin present in most erythrocytes. Hydroxyurea, which increases levels, widely available effective, especially children. Nevertheless, only cell-based gene therapy achieve a 'curative' threshold.

Язык: Английский

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