48 results about "Ligase activity" patented technology

Methods of screening for modulators of TRIM24 (also known as TIF1-ALPHA) expression and / or biological activity are described. In particular, methods of screening of screening for modulators of TRIM24 E3 ligase activity, and specifically an E3 ligase activity directed at p53 as the target polypeptide are also described. Modulators of TRIM24 expression and activity are provided and their use in treatment of cancer, particularly in breast, colon, prostate, renal cancers and in acute lymphoblastic leukaemia. Suitable modulators of TRIM24 expression include siRNA and shRNA and can be used in the treatment of cancer and for targeting cancer stem cells.

The present invention relates to a method for detecting the post-translational modification of a target protein by a post translational modifier polypeptide molecule; a method for screening a candidate protein for E3 ligase activity; a method of screening a test compound for the ability to regulate the post-translational modification of a target protein by a post-translational modifier polypeptide molecule; and a method for the large-scale detection of candidate target proteins of post-translational modification by a modifier polypeptide molecule. The present invention also relates to a kit for determining whether a test protein is post-translationally modified by a modifier polypeptide molecule; a kit for screening a test compound for the ability to regulate the post-translational modification of a target protein by a post-translational modifier peptide molecule, and another kit for determining whether a test protein is post-translationally modified by a modifier polypeptide molecule.

The invention provides methods of preventing or treating a condition associated with DNA damage in an animal comprising the administration of a substance that interferes with the activity of the CUL4A ubiquitin ligase. The invention also provides a substance that interferes with the activity of CUL4A, as well as compositions comprising the interfering substance and a carrier. The substance of the invention preferably enhances nucleotide excision repair activity in an animal. The invention further provides methods of identifying substances that negatively or positively modulate the expression and / or activity of CUL4A.

Provided herein are compositions, systems, and methods employing hyper-thermostable lysine-mutant ssDNA / RNA ligases that possesses both ssRNA ligase and ssDNA ligase activity. In certain embodiments,such hyper-thermostable lysine-mutant ssDNA / RNA ligases are used to ligate an first single stranded nucleic acid sequence with a 5' adenylated end to a second single stranded nucleic acid sequence (e.g., at a temperature of at least 75 DEG C) to form a ligated nucleic acid sequence. In further embodiments, the ligated nucleic acid sequence is sequenced.

The present invention describes methods of molecular diagnosis of a concrete form of a-synucleinopathy, the dementia with Lewy bodies (DLB), associated to the levels of ubiquitin carboxy-terminal hydrolase L1 (UCH-1) or the alteration of its ubiquityl-ligase activity. It also refers to the use of compounds that permit the modification of the UCH-L1 levels or of the enzymatic activity of UCH-L1. This invention has application in the diagnosis and treatment of patients suffering from DLB.

The present invention generally relates to compounds and compositions useful for the modulation of ligase activity. The invention further relates to Compounds of the Invention, compositions thereof, and methods for treating or preventing cancer, a neoplastic disorder, acute or chronic renal failure, an inflammatory disorder, an immune disorder, a cardiovascular disease, an effect of aging or an infectious disease comprising administering an effective amount of a Compound of the Invention. The invention further relates to the use of a Compound of the Invention as a preservative of a cell, blood, tissue or an organ or as an agent to modulate stem cells.

The present invention relates to methods that utilize multi-component nucleic acid complex (MNA complex) cascades. The MNA complexes may have cleavage or ligase activity. Further, the invention provides cascades which may include one or more DNAzymes. The invention also provides methods which use these cascades for the identification, detection and quantification of targets.

The present invention describes methods of molecular diagnosis of a concrete form of a-synucleinopathy, the dementia with Lewy bodies (DLB), associated to the levels of ubiquitin carboxy-terminal hydrolase L1 (UCH-L1) or the alteration of its ubiquityl-ligase activity. It also refers to the use of compounds that permit the modification of the UCH-L1 levels or of the enzymatic activity of UCH-L1. This invention has application in the diagnosis and treatment of patients suffering from DLB.

The present invention generally relates to compounds and compositions useful for the modulation of ligase activity. The invention further relates to Compounds of the Invention, compositions thereof, and methods for treating or preventing cancer, a neoplastic disorder, acute or chronic renal failure, an inflammatory disorder, an immune disorder, a cardiovascular disease, an effect of aging or an infectious disease comprising administering an effective amount of a Compound of the Invention. The invention further relates to the use of a Compound of the Invention as a preservative of a cell, blood, tissue or an organ or as an agent to modulate stem cells.

Methods of screening for modulators of TRIM24 (also known as TIF1-ALPHA) expression and / or biological activity are described. In particular, methods of screening of screening for modulators of TRIM24 E3 ligase activity, and specifically an E3 ligase activity directed at p53 as the target polypeptide are also described. Modulators of TRIM24 expression and activity are provided and their use in treatment of cancer, particularly in breast, colon, prostate, renal cancers and in acute lymphoblastic leukaemia. Suitable modulators of TRIM24 expression include siRNA and shRNA and can be used in the treatment of cancer and for targeting cancer stem cells.

The present invention generally relates to compounds and compositions useful for the modulation of ligase activity. The invention further relates to Compounds of the Invention, compositions thereof, and methods for treating or preventing cancer, a neoplastic disorder, acute or chronic renal failure, an inflammatory disorder, an immune disorder, a cardiovascular disease, an effect of aging or an infectious disease comprising administering an effective amount of a Compound of the Invention. The invention further relates to the use of a Compound of the Invention as a preservative of a cell, blood, tissue or an organ or as an agent to modulate stem cells.

A method of detecting a ligase expressing micro-organism in a sample comprises steps of treating the sample under conditions that inhibit the activity of ATP-dependent ligase from mammalian cells but which do not inhibit the activity of the microbial ligases, contacting the sample or a portion of the sample with a nucleic acid molecule which acts as a substrate for ligase activity in the sample, incubating the thus contacted sample under conditions suitable for ligase activity; and specifically determining the presence and / or the amount of a ligated nucleic acid molecule resulting from the action of the ligase on the substrate nucleic acid molecule to indicate the presence of the ligase expressing micro-organism. The micro-organism may be a fungus or a bacterium or both. High pH conditions may be employed to inactivate mammalian ligases. Related kits are described.

The present invention relates to compositions and methods for disease treatment and prevention through administration of a live attenuated composition. In particular, the present invention provides compositions and methods for the treatment and prevention of urinary tract infection by administration of a live attenuated compositions lacking O-antigen ligase activity.

The present invention provides a novel double-stranded DNA peptide ligase dDPlaseI, which can be used to catalyze the covalent connection of a specific double-stranded DNA and a specific polypeptide, and its enzymatic properties and use methods. The amino acid sequence of the dDPlaseI enzyme is shown in SEQ ID NO: 6, and the corresponding gene coding sequence is shown in SEQ ID NO: 5; The 5'-terminal deoxyribonucleotide A is a specific DNA double-strand in the phosphorylated state and the C-terminal start is a specific polypeptide chain with 7 peptides of N'‑MANCEHL‑C', and catalyzes the covalent bond between the two Together; double-stranded DNA and polypeptide ligation products have characteristic absorption peaks at 360nm wavelength, which can be used for the determination of dDPlaseI ligase activity. dDPlaseI ligase buffer was composed of 450 mM Tris-HCl, 100 mM Mg, pH 7.8 2+ , 80mM NaCl, 20mM ATP and 8mM Triton X-100, the optimal reaction temperature range is 35℃-45℃, and the ligation reaction time is 3min-10min. The novel double-stranded DNA polypeptide ligase provided by the present invention can be used as a tool enzyme for genetic engineering and gene analysis.

The present invention utilizes three families of bacterial enzymes, which play a key role in mycothiol biosynthesis. The three families are bacterial cysteine:glucosaminyl inositol ligases (MshC) with catalytic ligase activity for ligation of glucosaminyl inositol and cysteine, bacterial acetyl-CoA:Cys-GlcN-Ins acetyltransferases (MshD) with catalytic activity for addition of an acetyl group to Cys-GlcN-Ins and bacterial MshA glycosyltransferase with catalytic activity for production of GlcNAc-Ins. The invention provides methods for using the mycothiol biosynthesis ligases, acetyltransferases or glycosyltransferases in drug screening assays to determine compounds that inhibit activity. The invention provides for treatment of actinomycete infections in mammals using antibiotics that inhibit production or activity of the enzymes of mycothiol biosynthesis, in particular MshC, MshD or MshA, and thereby reduce the production of mycothiol and the virulence of the infecting bacteria. Additionally, the invention provides a live mutant with a genome containing a modification in an endogenous enzyme of mycothiol biosynthesis gene. The invention also provides an expression vector comprising polynucleotides of mshA, mshB, mshC and mshD.

The present invention relates to a method for screening and identifying compounds that modulate the activity of one or more components in the tRNA splicing pathway. In particular the invention relates to a method for screening and identifying compounds that modulate the activity tRNA splicing endonuclease and / or tRNA splicing ligase. The invention provides assays for the identification of compounds that inhibit animalia tRNA splicing endonuclease and / or animalia tRNA splicing ligase. The invention also provides assays for the identification of compounds that inhibit fungal tRNA splicing endonuclease and / or fungal tRNA splicing ligase. The methods of the present invention provide a simple, sensitive assay for high-throughput screening of libraries of compounds to identify pharmaceutical leads useful for treating and / or preventing cancer and / or fungal infections.

More than 2% of adults harbor a pancreatic cyst, a subset of which progress to invasive lesions with lethal consequences. To assess the genomic landscapes of neoplastic cysts of the pancreas, we determined the exomic sequences of DNA from the neoplastic epithelium of eight surgically resected cysts of each of the major neoplastic cyst types: serous cystadenomas (SCAs), intraductal papillary mucinous neoplasms (IPMNs), mucinous cystic neoplasms (MCNs) and solid pseudo-papillary neoplasms (SPNs). SPNs are low-grade malignancies, and IPMNs and MCNs, but not SCAs, have the capacity to progress to cancer. We found that SCAs, IPMNs, MCNs, and SPNs contained 10=4.6, 27=12, 16=7.6, and 2.9=2.1 somatic mutations per tumor, respectively. Among the mutations identified, E3 ubiquitin ligase components were of particular note. Four of the eight SCAs contained mutations of VHL, a key component of the VHL ubiquitin ligase complex that has previously been associated both with renal cell carcinomas, SCAs, and other neoplasms. Six of the eight IPMNs and three of the eight MCNs harbored mutations of RNF43, a gene coding for a protein with intrinsic E3 ubiquitin ligase activity that has not previously been found to be genetically altered in any human cancer. The preponderance of inactivating mutations in RNF43 unequivocally establish it as a suppressor of both IPMNs and MCNs. SPNs contained remarkably few genetic alterations, but always contained mutations of CTNNB1, previously demonstrated to inhibit degradation of the encoded protein (β-catenin) by E3 ubiquitin ligases. These results highlight the essential role of ubiquitin ligases in these neoplasms and have important implications for the diagnosis and treatment of patients with cystic tumors.

The invention discloses a method for easily and rapidly evaluating the efficiency of a molecular cloning system. A DNA fragment is designed, at least two multiple cloning sites are arranged on the DNAfragment, restriction enzyme digestion sites of to-be-assayed restriction endonuclease are provided in the multiple cloning sites, and the restriction enzyme digestion sites only exist in the multiple cloning sites; sticky ends which are produced in the different multiple cloning sites by the same endonuclease can be complementarily connected; for the DNA fragment undergoes restriction enzyme digestion, electrophoresis is carried out after the restriction enzyme digestion product is connected by ligase, and the formed electrophoretic band is analyzed: if the electrophoretic band exists as a large fragment or a single complete vector fragment, then the endonuclease has lower or no activity; and if the electrophoretic band exists as a small fragment, then the ligase has lower or no activity. The provided method for evaluating the efficiency of a molecular cloning system can easily and rapidly test the activity of a main ingredient (endonuclease or ligase) in a restriction enzyme digestion connecting system.