50 results about "Deubiquitination" patented technology

Compositions, including antibodies, polypeptides, and organic molecules, kits, and methods for probing molecular interactions (e.g., deubiquination, ubiquination and kinase activity), e.g., using resonance energy transfer (RET) are provided.

The present invention discloses an Ub-Nanoluc reporter gene system and an Ub-Ub-GS-Nanoluc reporter gene system, and applications of the Ub-Nanoluc reporter gene system and the Ub-Ub-GS-Nanoluc reporter gene system in deubiquitination enzyme activity detection and deubiquitination enzyme inhibitor throughput-screening, wherein the Ub-Nanoluc reporter gene DNA sequence is represented by SEQ ID NO.1, and the Ub-Ub-GS-Nanoluc reporter gene DNA sequence is represented by SEQ ID NO.4. According to the present invention, the high sensitivity characteristic of luciferase Nanoluc is utilized to provide the method for deubiquitination enzyme activity detection and deubiquitination enzyme inhibitor throughput-screening by using the chemical luminescent reporter gene so as to provide the new action target point and the new ideal for screening and research of new drugs including anticancer drugs, and provide broad application prospects, great enormous benefits, and great social benefits.

The hypoxia inducible factor-1 (HIF-1) transcription factor is an important regulator of the cellular response to hypoxia. The activity of HIF-1 is regulated by the level of the HIF-1α subunit, HIF-1α, which is rapidly degraded under normoxic conditions by the ubiquitin-proteasome pathway. HIF-1α levels increase under hypoxic conditions. Many human cancers also show constitutively increased HIF-1α levels. PX-478 or S-2-amino-3-[4′-N,N,-bis(2-chloroethyl)amino]phenyl propionic acid N-oxide dihydrochloride, is a novel anticancer agent, and is capably of decreasing both constitutive and hypoxia induced HIF-1α protein levels and HIF-1 transactivation in vitro and in vivo. In method embodiments, the administration of PX-478 is independent of the pathways of HIF-1α regulation involving the von Hippel-Lindau protein and p53. PX-478 causes an increase in polyubiquitinated HIF-1α levels due to inhibition of HIF-1α deubiquitination. The levels of other proteins whose proteasomal breakdown is mediated by ubiquitination are not affected by PX-478. Deubiquitination is a novel pathway for the regulation of cellular HIF-1α levels and PX-478 is a specific inhibitor of the pathway. Therapeutic compounds for regulating cellular HIF-1α levels and methods of regulating cellular HIF-1α levels are herein provided.

Viruses having an impaired ability to deISGylate ISG15 conjugates, in particular, viral mutants comprising a mutation in the viral genome that reduces or eliminates the ability of the viral OTU domain-containing protein encoded by the viral genome to deISGylate ISG15 conjugates and/or deubiquitinate ubiquitinated proteins and/or deNeddylate Neddylated proteins are disclosed. Such viral mutants may be used in the formulation of immunogenic compositions for inducing an immune response and preventing, managing and/or treating a viral infection. Also disclosed are methods for identifying anti-viral compounds, in particular, methods of identifying compounds that reduce or inhibit the deISGylation activity and/or deubiquitination and/or deNeddylation activity of a viral OTU domain-containing protein. The compounds identified using such methods may be used as antiviral agents for the prevention, treatment and/or management of viral infections.

Disclosed are small molecule inhibitors of deubiquitinating enzymes (DUBs), and methods of using them. Certain compounds display a preference for specific ubiquitin specific proteases (USPs).

The invention relates to an application of a ubiquitination pathway related factor in regulating and controlling functions of a regulatory T cell. Specifically, the invention relates an application ofthe ubiquitination pathway related factor and an agonist or an antagonist thereof in preparing a preparation or a reagent for regulating FOXP3 or the regulatory T cell, wherein the ubiquitination related factor is selected from gene coding proteins USP44 and USP7 of a ubiquitination protein family. The novel regulatory factor provided by the invention, by regulating transcriptional regulatory activity of the FOXP3 on a target gene thereof, can regulate and control the regulatory T cell and the balance of an immune system.

The invention relates to a modified Bach1 gene and an application thereof to pluripotency, self-updating, proliferation, reprogramming and differentiation of a stem cell. Due to the modification of the Bach1 gene of a transcription factor, the BACH1 protein activity or expression of the cell is inhibited or increased, and furthermore, the cell interacts with a deubiquitinated protease Usp7, a pluripotent factor Nanog, Sox2 and/or Oct4, a PRC2 complex, a Wnt/beta-catenin signal channel and/or an Nodal/Smad2/3 signal channel, so that the self-updating, proliferation and/or differentiation of thestem cell are regulated.

The invention belongs to the technical field of deubiquitination enzyme fluorescence detection, and particularly relates to a preparation method of a nano fluorescent probe for detecting deubiquitination enzyme. The preparation method comprises following steps: dispersing mesoporous silicon nano particles into a dichloromethane solution containing phenanthroline-silicon, carrying out reactions toobtain mesoporous silicon-phenanthroline nano particles; mixing mesoporous silicon-phenanthroline nano particles and TbCl3.6H2O in ethanol, refluxing, adding 1,3-diphenyl-1,3-propanedione, carrying out reactions to obtain a nano fluorescent probe precursor; suspending the nano fluorescent probe precursor in a polyethyleneimine water solution to coat polyethyleneimine to obtain a polyethyleneiminemodified nano fluorescent probe precursor; dispersing the polyethyleneimine modified nano fluorescent probe precursor into the mixed solution, and stirring to obtain the nano fluorescent probe. The preparation process is simple, the period is short; and the prepared nano fluorescent probe has good sensitivity and selectivity for detection of UCH-L1.

The invention discloses an application of a USP9x inhibitor in preparation of a medicine for treating osteosarcoma. By discovering that the degradation of a proliferation-promoting factor SOX2 can beeffectively promoted by reducing the expression of USP9x or inhibiting the activity of the USP9x in the osteosarcoma, a new target is provided for clinically developing new chemotherapeutic medicinesfor the osteosarcoma; and meanwhile, the research also discovers that NGA can effectively inhibit the deubiquitination enzyme activity of the USP9x, thereby indirectly promoting the ubiquitination degradation of the SOX2, inhibiting the generation and development of tumors, and providing a new molecular structure for clinically discovering new medicines.

The invention discloses a function of a deubiquitination enzyme ZRANB1 for regulating lipid metabolism and application of the deubiquitination enzyme ZRANB1. The invention further discloses application of inhibiting ZRANB1 gene expression in preparing a product with at least one function of treating and/or preventing obesity, promoting white fat cell combustion, promoting slimming and losing weight and inhibiting accumulation/accumulation of subcutaneous and/or intra-abdominal white fat. Experiments prove that ZRANB1, as a newly discovered important fat metabolism regulation molecule, can significantly reduce the content of white fat of an organism after the protein level is reduced. The discovery of the ZRANB1 function provides an important scientific basis and application value for prevention of obesity diseases and treatment of slimming.

The invention discloses an application and a method of a USP10 gene and/or an Ascl1 gene in inducing transdifferentiation of fibroblasts into neuronal cells. The invention firstly discloses the application of the USP10 gene and/or the Ascl1 gene in inducing the fibroblast to be transdifferentiated into the neuronal cell or the application in preparing a product for inducing the fibroblast to be transdifferentiated into the neuronal cell. The invention further discloses a method for inducing transdifferentiation of fibroblasts into neuronal cells. The USP10 capable of improving the stability of the Ascl1 protein and deubiquitinating the Ascl1 is obtained by screening based on the characteristics of the Ascl1. The USP10 gene and the Ascl1 gene are applied to the process of converting fibroblasts into neuronal cells, the GABAergic neuronal cells can be efficiently obtained, the defect that the neuronal cells cannot be efficiently obtained is overcome, and the application has important significance on treatment of nerve-related diseases.

The invention provides a deubiquitination enzyme activity detection method based on fluorescence polarization. The method comprises the following steps: 1, preparing a deubiquitinating enzyme detection probe: preparing a fusion protein Ub-linker-Cys, connecting a BODIPY probe, and carrying out mass spectrometric detection; 2, detecting the activity of the deubiquitination enzyme: selecting the deubiquitination enzyme and detecting the activity of the deubiquitination enzyme in real time according to the influence of deubiquitination on the FP value of the probe. The detection method is based on detection of fluorescence polarization, the fluorescence polarization detects the proportional relation of the fluorescence intensity in the parallel direction and the fluorescence intensity in the vertical direction, the detection method is not easily interfered by the concentration of the fluorescence probe, the solution temperature, the pH value and other fluorescence signals, when the activity of the deubiquitination enzyme is measured, the steps are simple, the consumed time is short, and the usage amount of the probe and the deubiquitination enzyme is small. In addition, the probe prepared in the method can also be used for screening deubiquitination enzyme inhibitors, and is high in throughput.

The invention relates to a lysine site mutant of conjugated ubiquitin of mda-7/IL-24 and application thereof, belonging to the field of genetic engineering. In the invention, lysine codons of 63rd, 69th, 78th, 119th, 123rd, 136th, 179th, 189th, 203rd and 206th potential conjugated ubiquitin sites of the mda-7/IL-24 are mutated into arginine codons by utilizing a site-directed mutation PCR (Polymerase Chain Reaction) technology and cloned to pcDNA3.1(+) plasmids; after the mutant plasmids are transfected with cervical carcinoma Hela cells and hepatoma carcinoma HepG2 cells, the expressions of mda-7/IL-24 protein and ubiquitination mda-7/IL-24 protein are respectively detected. A result shows that the quantity of the mda-7/IL-24 protein expressed by a 123rd lysine mutant of the mda-7/IL-24 is remarkably increased, and the quantity of the ubiquitination mda-7/IL-24 protein is remarkably decreased. The invention proves that the 123rd lysine of the mda-7/IL-24 is an ubiquitination site, and mda-7/IL-24 mutants generated by the mutant plasmids are difficult to be degraded by the in-vivo ubiquitin, thereby enhancing the expression and the action of the mda-7/IL-24 in tumour cells.