30 results about "S-methyltransferase activity" patented technology

Methods for amplifying a nucleic acid sample while preserving the methylation status of cytosines are disclosed. In some aspects the amplified methylated sample is modified by methylation sensitive modification and analyzed by hybridization to an array to identify cytosines that were methylated in the starting material and cytosines that were not methylated in the starting material. Methods for detecting methylation status are also disclosed. In one embodiment a DNA methyltransferase activity is included in the amplification reaction and this activity methylates the newly synthesized DNA using the methylated genomic template strand as a guide.

The present invention features a method for determining the methyltransferase activity of a polypeptide and screening for modulators of methyltransferase activity, more particularly for modulators of the methylation of retinoblastoma by SMYD3. The invention further provides a method or pharmaceutical composition for prevention or treating of colorectal cancer, hepatocellular carcinoma, bladder cancer and / or breast cancer using a modulator so identified. N-terminal truncated forms of SMYD3 (alias ZNFN3A1) have higher methylating activity. Lys 824 is a preferred methylation site on the RB1 protein for SMYD3.

The invention discloses a method for detecting the activity of DNA methylase and DNA methyltranseferase based on restriction endonuclease Hpa pi and exonuclease III. An unlabeled DNA probe is prepared and the sequence of the probe comprises a cleavage site of the restriction endonuclease and a methylated CpG site. The purpose of detecting the activity of the methylase and methyltranseferase is reached by steps of hybridizing the unlabeled DNA probe with the target DNA, carrying out methylation treatment by the methylase, carrying out the specific cleavage of the restriction endonuclease Hpa pi and enabling exonuclease III having the activity of 3'->5' exonuclease to act on the double-stranded DNA and then adding fluorescence signal molecules thiazole orange which having different signals to single-stranded DNA and double-stranded DNA. According to the method, since no complex material or labeling of DNA probes need to be prepared, the defects of high detection cost, cumbersome operations and poor reproducibility caused by the preparation of the material and labeling of DNA probes are avoided. The method disclosed by the invention has the advantages of low cost and high sensitivity and is fast and simple.

The invention features a method for determining methyl transferase activity of a polypeptide and screening for modulators of methyl transferase activity. The invention further provides a method or pharmaceutical composition for prevention or treating of colorectal cancer or hepatocellular carcinoma using the modulator.

The described invention provides methods for modulating gene expression of a gene related to proliferation of a population of tumor cells. The method entails administering a therapeutic amount of a therapeutic compound to a cell, a tissue, or a mammal, wherein the therapeutic amount of the therapeutic compound is effective to suppress methyltransferase activity of a protein arginine methyltransferase. Modulation of the protein arginine methyltransferase activity in turn modulates methylation of a target protein that affects gene expression of the gene, and may suppress the proliferation of the population of tumor cells.

Murine and human Suv39h2 polypeptide and DNA molecules encoding them. Suv39h2 is a novel member of the Suv3-9 gene family. Suv39h2 is a novel component of meiotic higher order chromatin. It has histone methyltransferase activity and is required, in combination with Suv39h1, for male gametogenesis. Suv39h2 can be used in screening methods to identify modulators of its methyltransferase activity, which are useful in cancer therapy and for male contraception.

Methods are described for measuring the amount of a methylation TPMT enzyme product in a sample. More specifically, mass spectrometric methods are described for detecting and quantifying 6-MMP or isotopically labeled 6-MMP in a test sample utilizing mass spectrometric techniques and for using such methods to determine the activity of TPMT enzyme that is present in a sample.

Methods are disclosed for determining the methylation state of DNA samples using melt analysis including high resolution melt analysis. Methods are also provided for determining methyltransferase activity using melt analysis including high resolution melt analysis. Additionally, kits of parts are provided.

The invention discloses a method for measuring the activity of transmethylase in real time and a kit. The method comprises two reactions carried out at the same time in a reaction system; according to the first reaction, in a buffer system where it is guaranteed that S-ademetionine is adopted as a methyl donor and the transmethylase has the biological activity, a sample containing the transmethylase (MT), the methyl donor and a corresponding substrate are added, or the S-ademetionine methyl donor and the corresponding substrate are directly added into the liquid sample containing the transmethylase, and a reaction is carried out so that an S-adellosyl homocysteine (SAH) product can be generated, wherein the substrate is receptor matter of methyl; according to the second reaction, an immunological method is adopted for detecting the content of SAH to determine the activity of the transmethylase in the reaction system. The biochemical reaction of MT catalysis and the immunoreactions for measuring the product SAH are carried out at the same time, the two processes are organically combined, and great significance is especially achieved on accurate measurement of SAH with extremely unstable molecularity.

The present invention relates generally to a genetic sequence encoding a polypeptide having methyltransferase activity and the use of the genetic sequence and / or the polypeptide to modify one or more phenotypic characteristics of a plant. More particularly, the methyltransferase of the present invention acts on flavonoids, preferably wherein the flavonoid is an anthocyanin. Even more particularly, the present invention relates to a polypeptide having S-adenosyl-L-methionine:anthocyanin 3′-O-methyl-transferase or S-adenosyl-L-methionine:anthocyanin 3′,5′-O-methyltransferase activity. The present invention still further provides a genetic sequence encoding a polypeptide having methyltransferase activity derived from Petunia, Torenia Fuchsia or Plumbago or botanically related plants. The instant invention further relates to antisense and sense molecules corresponding to all or part of the subject genetic sequence as well as genetically modified plants as well as cut flowers, parts, extracts and reproductive tissue from such plants.

The present invention provides an isolated nucleic acid molecule encoding a PFM / SET polypeptide. Also provided is an isolated nucleic acid molecule encoding a functional fragment of a PFM / SET polypeptide that contains a PR, SET, PRAZ or PKZL domain of a PFM / SET polypeptide of the invention. Further provided by the invention are PFM / SET polypeptides, and functional fragments thereof that contain a PR, SET, PRAZ or PKZL domain of a PFM / SET polypeptide. The invention also provides PFM / SET antibodies, PFM / SET modulatory compounds, and related methods. The molecules of the invention can be used in methods of screening for a compound that modulates PFM / SET polypeptide histone methyltransferase activity and to modulate cell proliferation to prevent or treat proliferative disorders, including cancer. Additionally, the molecules and methods of the invention can be used to diagnose and prognose proliferative disorders.

The present invention relates to genes associated with the tocopherol biosynthesis pathway. More particularly, the present invention provides and includes nucleic acid molecules, proteins, and antibodies associated with genes that encode polypeptides that have methyltransferase activity. The present invention also provides methods for utilizing such agents, for example in gene isolation, gene analysis and the production of transgenic plants. Moreover, the present invention includes transgenic plants modified to express the aforementioned polypeptides. In addition, the present invention includes methods for the production of products from the tocopherol biosynthesis pathway.

The current disclosure reveals a complex regulatory pattern between miR-31 and AR, indicating that miR-31 plays a key role in prostate cancer development and progression. Another aspect of the current disclosure shows that miR-31 directly targets and destabilizes AR mRNA through interaction with the AR mRNA coding sequence showing that miR-31, or a fragment thereof has the ability to act as a novel therapeutic agent in treating cancer. The current disclosure also shows that AR indirectly represses miR-31 expression by binding to the miR-31 promoter region and modulating methyltransferase activity. Another aspect of the current disclosure shows that miR-31 indirectly modulates AR activity by modulating regulators of cell cycle progression. The disclosure further provides an isolated nucleic acid that modulates the activity of the androgen receptor in a cell. The disclosure further provides a method of treating a prostate cancer in a subject, by administering to the subject an effective amount of an agent that modulates the activity or levels of miR-31.

The present invention relates to coupled enzyme assays. In particular, the present invention provides a coupled fluorescent assay for detection of S-adenosylmethionine (AdoMet)-dependent methyltransferase activity.

The present invention relates to genes associated with the tocopherol biosynthesis pathway. More particularly, the present invention provides and includes nucleic acid molecules, proteins, and antibodies associated with genes that encode polypeptides that have methyltransferase activity. The present invention also provides methods for utilizing such agents, for example in gene isolation, gene analysis and the production of transgenic plants. Moreover, the present invention includes transgenic plants modified to express the aforementioned polypeptides. In addition, the present invention includes methods for the production of products from the tocopherol biosynthesis pathway.

The invention discloses a method for quantitative determination of catechol-o-methyl transferase activity and a use thereof and belongs to the technical field of biological medicines. The method utilizes 4-acetoxy-7,8-dihydroxycoumarin as a COMT enzyme specific probe substrate. The substrate can be specifically catalyzed through the COMT enzyme to produce a 8-methoxy product, the 8-methoxy product can produce a fluorescence signal at wavelength of 520nm and through detection of fluorescence intensity change, COMT activity is detected quantificationally. The method can be used for quantitative evaluation of activity of COMT enzymes in biological samples with different sources and can also be used for external rapid screening of COMT inhibitors and assessment of an inhibition capability. The method can be used for quantitative assessment of real activity of COMT enzymes in various external biological samples, realizes inhibitor rapid screening and inhibition capability quantitative evaluation and can be used for evaluating catalytic activity of different types of COMTs and COMT mutants with different amino acid sequences and further evaluating a catechol drug metabolism capability. The method has the advantages of high selectivity, low price, acquisition easiness and high sensitivity.

Disclosed are novel methyltransferase assay methods, comprising: including, in a reaction mixture for a methyltransferase activity, a purified or recombinant adenosine nucleosidase activity that catalyses release of an adenine or adenine derivative moiety from a transmethylation product, and a purified or recombinant adenine deaminase activity that catalyses deamination of the released moiety to hypoxanthine or respective derivative and ammonia, wherein the methyltransferase activity is rate-limiting; and determining the methyltransferase activity by spectrophotometric or chromatographic monitoring of the coupled deamination reaction products, or of subsequent enzymatic or chemical reactions coupled thereto. Coupled oxidation of the hypoxanthine to uric acid and hydrogen peroxide is optionally affected using purified or recombinant xanthine oxidase, wherein the methyltransferase activity is rate-limiting, and wherein determining the methyltransferase activity comprises monitoring of the coupled oxidation reaction. Variations are disclosed comprises monitoring of reaction products (e.g., to detect NH3, Hypoxanthine, H2O2, and Uric Acid).

The invention discloses a method for detecting the activity of DNA methylase and DNA methyltranseferase based on restriction endonuclease Hpa pi and exonuclease III. An unlabeled DNA probe is prepared and the sequence of the probe comprises a cleavage site of the restriction endonuclease and a methylated CpG site. The purpose of detecting the activity of the methylase and methyltranseferase is reached by steps of hybridizing the unlabeled DNA probe with the target DNA, carrying out methylation treatment by the methylase, carrying out the specific cleavage of the restriction endonuclease Hpa pi and enabling exonuclease III having the activity of 3'->5' exonuclease to act on the double-stranded DNA and then adding fluorescence signal molecules thiazole orange which having different signals to single-stranded DNA and double-stranded DNA. According to the method, since no complex material or labeling of DNA probes need to be prepared, the defects of high detection cost, cumbersome operations and poor reproducibility caused by the preparation of the material and labeling of DNA probes are avoided. The method disclosed by the invention has the advantages of low cost and high sensitivity and is fast and simple.

The invention relates to a method for detecting the activity of catechol-oxygen-methyltransferase (COMT) in blood and belongs to the technical field of biological medicine. According to the invention,3-benzothiazole-7,8-dihydroxycoumarin (3-BTD) with high chemical stability is selected as the fluorescence substrate of COMT enzyme by virtue of the advantages of a high performance liquid chromatography and fluorescence detection combined instrument on biological matrix interference resistance and detection sensitivity; meanwhile, a high-efficiency and high-sensitivity measuring method capable of quantitatively analyzing the activity of a trace amount of COMT in the blood is developed by combining a high performance liquid chromatography-fluorescence detection technology. A fluorescence probe substrate adopted by the method can be obtained through chemical synthesis, the synthesis process is simple and practical, and the performance is stable. The method has the advantages of high sensitivity, high accuracy degree, high biological matrix interference resistance, high repeatability and the like, can be applied to detection on the activity of the COMT enzyme in the blood, and also canbe applied to quantitative measurement on the activity of the COMT enzyme in various mammal-derived cells or tissue preparation objects as well as screening and evaluation of a COMT inducer / activator.

Juvenile hormone acid methyltransferase gene was cloned using a cDNA derived from the corpora allata of Bombyx mori by differential display methods. A recombinant protein expressed in Escherichia coli, which was transformed with a vector DNA incorporating the gene, was found to have juvenile hormone acid methyltransferase activity. Based on amino acid sequence homology, juvenile hormone acid methyltransferase genes were found from Drosophila melanogaster, Anopheles gambiae, Spodoptera litura, and Helicoverpa armigera. The proteins encoded by the juvenile hormone acid methyltransferase genes derived from Drosophila melanogaster, Spodoptera litura, and Helicoverpa armigera were found to have juvenile hormone acid methyltransferase activities.

The invention provides a cascade rolling-ring amplification strategy mediated with a dumbbell molecular probe under protection of a target object and applied to sensitiveness detection of DNA transmethylase activity. Based on the cascade rolling-ring amplification strategy mediated with the dumbbell molecular probe under protection of the target object, a novel fluorescence detection system for DNA transmethylase activity analysis is developed. The detection system has excellent specificity and sensitivity, the detection limit is as low as 0.0024 U / mL, and potential application value is exhibited in quantitative monitoring of transmethylase activity and screening of anticarcinogen.

The invention features a method for determining methyl transferase activity of a polypeptide and screening for modulators of methyl transferase activity. The invention further provides a method or pharmaceutical composition for prevention or treating of colorectal cancer or hepatocellular carcinoma using the modulator.

The invention relates to an attenuated non-segmented negative-sense RNA virus characterized by at least one mutation in the L gene wherein the mutation reduces viral replication, the methods of manufacturing and methods of use.