Small molecule control of intracellular protein levels allows temporal and dose-dependent regulation of protein function. Recently, we developed a method to degrade proteins fused to a mutant dehalogenase (HaloTag2) using small molecule hydrophobic tags (HyTs). Here, we introduce a complementary method to stabilize the same HaloTag2 fusion proteins, resulting in a unified system allowing bidirectional control of cellular protein levels in a temporal and dose-dependent manner. From a small molecule screen, we identified N-(3,5-dichloro-2-ethoxybenzyl)-2H-tetrazol-5-amine as a nanomolar HALoTag2 Stabilizer (HALTS1) that reduces the Hsp70:HaloTag2 interaction, thereby preventing HaloTag2 ubiquitination. Finally, we demonstrate the utility of the HyT/HALTS system in probing the physiological role of therapeutic targets by modulating HaloTag2-fused oncogenic H-Ras, which resulted in either the cessation (HyT) or acceleration (HALTS) of cellular transformation. In sum, we present a general platform to study protein function, whereby any protein of interest fused to HaloTag2 can be either degraded 10-fold or stabilized 5-fold using two corresponding compounds.
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4113957 | PMC |
| http://dx.doi.org/10.1021/cb400569k | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Anatomy and transcriptomics of the common jingle shell (Bivalvia, Anomiidae) support a sensory function for bivalve tentacles.
Sci Rep
December 2024
Department of Zoology, University of São Paulo, São Paulo, SP, Brazil.
Animals have evolved numerous mechanisms to perceive and interact with the environment that can be translated into different sensory modalities. However, the genomic and phenotypic features that support sensory functions remain enigmatic for many invertebrates, such as bivalves, an ecologically and economically important taxonomic group. No repertoire of sensory genes has been characterized in bivalves, representing a significant knowledge gap in molluscan sensory biology.
View Article and Find Full Text PDFPharmacological inactivation of a non-canonical gp130 signaling arm attenuates chronic systemic inflammation and multimorbidity induced by a high-fat diet.
Sci Rep
December 2024
Department of Orthopaedic Surgery, Keck School of Medicine, Stem Cell Research and Regenerative Medicine, University of Southern California, Los Angeles, CA, USA.
Interleukin-6 (IL-6) is a major pro-inflammatory cytokine that demonstrates a robust correlation with age and body mass index (BMI) as part of the senescence-associated secretory phenotype. IL-6 cytokines also play a crucial role in metabolic homeostasis and regenerative processes primarily via the canonical STAT3 pathway. Thus, selective modulation of IL-6 signaling may offer a unique opportunity for therapeutic interventions.
View Article and Find Full Text PDFA new aguanidine-based bis Schiff base for highly selective Al recognition, BSA binding studies and theoretical calculations.
Spectrochim Acta A Mol Biomol Spectrosc
December 2024
School of Agriculture and Bioengineering, Heze University, Heze 274500, China. Electronic address:
Herin, the successful synthesis of a bis Schiff base (L) has been achieved using 2-hydroxy-1-naphthaldehyde and 1,3-diaminoguanidine as raw materials, which was further characterized by infrared spectroscopy, mass spectrometry, and nuclear magnetic resonance hydrogen spectrum. Moreover, spectroscopic experiments demonstrated that the probe L showed good selectivity and visual detectability for Al. Its detection limit (DL) is 2.
View Article and Find Full Text PDFThe role of biodegradable plastics in lignite anaerobic digestion: Changes of organics transformation and metabolic pathway.
Bioresour Technol
December 2024
School of Energy Science and Engineering, Henan Polytechnic University, Jiaozuo 454000, China. Electronic address:
Biodegradable plastics (BPs) and lignite, both rich in organic matter, present significant challenges for efficient conversion into clean energy. This study examined the anaerobic co-digestion of BPs and lignite under controlled laboratory conditions. The results demonstrated that the co-digestion of polylactic acid (PLA) and lignite (at a 1:2 mass ratio, with 5 g PLA and 10 g lignite as the model system) rapidly acclimated to the anaerobic environment, enhancing cumulative biogas production by 57 % compared to the mono-digestion of lignite alone.
View Article and Find Full Text PDFRecent advances in spatiotemporal control of the CRISPR/Cas9 system.
Colloids Surf B Biointerfaces
December 2024
School of Life Sciences, State Key Laboratory of Biocatalysis and Enzyme Engineering, Hubei University, Wuhan, Hubei 430042, China. Electronic address:
The CRISPR/Cas9 gene-editing technology, derived from the adaptive immune mechanisms of bacteria, has demonstrated remarkable advantages in fields such as gene function research and the treatment of genetic diseases due to its simplicity in design, precise targeting, and ease of use. Despite challenges such as off-target effects and cytotoxicity, effective spatiotemporal control strategies have been achieved for the CRISPR/Cas9 system through precise regulation of Cas9 protein activity as well as engineering of guide RNAs (gRNAs). This review provides a comprehensive analysis of the core components and functional mechanisms underlying the CRISPR/Cas9 system, highlights recent advancements in spatiotemporal control strategies, and discusses future directions for development.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!