S‐Adenosylmetliionine and methylation

Chiang, Petek K.; Gordon, Richard K.; Tal, Jacov; Zeng, Guo-hua; Doctor, Bhupendra P.; Pardhasaradhi, K.; McCann, Peter P.

doi:10.1096/fasebj.10.4.8647346

Cited by 810 publications

(571 citation statements)

References 86 publications

(77 reference statements)

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“…SAM is an important metabolite as it serves as a major donor of methyl groups in numerous reactions involving methylation of DNA, RNA, proteins and metabolites [4][5][6] . Thus, in addition to its essential role in polyamine synthesis, AdoMetDC may also influence methylation reactions by affecting SAM availability.…”

Section: Adometdc Catalyses the Decarboxylation Of S-adenosylmethionimentioning

confidence: 99%

AMD1 mRNA employs ribosome stalling as a mechanism for molecular memory formation

Yordanova

Loughran

Zhdanov

et al. 2018

Nature

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Section: Adometdc Catalyses the Decarboxylation Of S-adenosylmethionimentioning

confidence: 99%

AMD1 mRNA employs ribosome stalling as a mechanism for molecular memory formation

Yordanova

Loughran

Zhdanov

et al. 2018

Nature

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“…Small organic molecules and peptides and most types of cellular macromolecules, including DNA, RNA, protein, and lipids, can serve as substrates for methylation (for detailed reviews, see Refs. [2][3][4][5][6][7]). …”

Section: Hhmi Author Manuscriptmentioning

confidence: 99%

“…The vast majority use Sadenosylmethionine (AdoMet) 1 as the methyl donor [1,2]. AdoMet is the second most …”

Section: Introductionmentioning

confidence: 99%

A universal competitive fluorescence polarization activity assay for S-adenosylmethionine utilizing methyltransferases

Graves

Zhang

Scott

2008

Analytical Biochemistry

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A high-throughput, competitive fluorescence polarization immunoassay has been developed for the detection of methyltransferase activity. The assay was designed to detect Sadenosylhomocysteine (AdoHcy), a product of all S-adenosylmethionine (AdoMet)-utilizing methyltransferase reactions. We employed commercially available anti-AdoHcy antibody and fluorescein-AdoHcy conjugate tracer to measure AdoHcy generated as a result of methyltransferase activity. AdoHcy competes with tracer in the antibody/tracer complex. The release of tracer results in a decrease in fluorescence polarization. Under optimized conditions, AdoHcy and AdoMet titrations demonstrated that the antibody had more than a 150-fold preference for binding AdoHcy relative to AdoMet. Mock methyltransferase reactions using both AdoHcy and AdoMet indicated that the assay tolerated 1 to 3 μM AdoMet. The limit of detection was approximately 5 nM (0.15 pmol) AdoHcy in the presence of 3 μM AdoMet. To validate the assay's ability to quantitate methyltransferase activity, the methyltransferase catechol-Omethyltransferase (COMT) and a known selective inhibitor of COMT activity were used in proofof-principle experiments. A time-and enzyme concentration-dependent decrease in fluorescence polarization was observed in the COMT assay that was developed. The IC 50 value obtained using a selective COMT inhibitor was consistent with previously published data. Thus, this sensitive and homogeneous assay is amenable for screening compounds for inhibitors of methyltransferase activity. KeywordsFluorescence polarization; Methyltransferase; S-Adenosylhomocysteine; S-Adenosylmethionine; Catechol-O-methyltransferase Methyltransferases are a diverse class of enzymes that catalyze the transfer of a single methyl group from a methyl donor molecule to a methyl acceptor. The vast majority use Sadenosylmethionine (AdoMet) 1 as the methyl donor [1,2]. AdoMet is the second most HHMI Author Manuscript HHMI Author Manuscript HHMI Author Manuscriptwidely used substrate molecule in the cell after ATP. Methylation is an essential and highly choreographed regulatory metabolic process. Small organic molecules and peptides and most types of cellular macromolecules, including DNA, RNA, protein, and lipids, can serve as substrates for methylation (for detailed reviews, see Refs. [2][3][4][5][6][7]).A rapidly growing area of methyltransferase research focuses on methylation of histone tails by histone methyltransferases (HMTs) (for reviews, see Refs. [8,9]). Histone methylation is an epigenetic modification that has a role in formation of heterochromatin, X chromosome inactivation, and modification of gene expression patterns. Histone methylation occurs on arginine and lysine residues and is catalyzed by methyltransferases belonging to three distinct families of proteins: the protein arginine N-methyltransferase 1 (PRMT1) family, the SET-domain-containing protein family, and the non-SET-domain proteins DOT1/DOT1L. This histone methylation alters chromatin structure and, thus, regulates...

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“…SAM is an important metabolic intermediate in all living cells as a donor of methyl groups in transmethylation reactions (2,3) and a precursor molecule in aminopropylation and transulfuration pathways (4). SAM also regulates the activities of various enzymes.…”

Section: Introductionmentioning

confidence: 99%

Expression of Secreted His-Tagged S-adenosylmethionine Synthetase in the Methylotrophic Yeast Pichia pastoris and Its Characterization, One-Step Purification, and Immobilization

Luo¹,

Yuan²,

Luo³

et al. 2008

Biotechnol. Prog.

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S-Adenosylmethionine synthetase (SAM synthetase) catalyzes the synthesis of S-adenosylmethionine (SAM), which plays an important role in cellular functions such as methylation, sulfuration, and polyamine synthesis. To develop a simple and effective way to enzymatically synthesize and produce SAM, a soluble form of SAM synthetase encoded by SAM2 from Saccharomyces cereVisiae was successfully produced at high level (∼200 mg/L) by the recombinant methylotrophic yeast Pichia pastoris. The secreted His 6 -tagged SAM synthetase was purified in a single chromatography step with a yield of approximately 82% for the total activity. The specific activity of the purified synthetase was 23.84 U/mg. The recombinant SAM synthetase could be a kind of allosteric enzyme with negative regulation. The enzyme functioned optimally at a temperature of 35°C and pH 8.5. The stability of the recombinant synthetase and the effectiveness of different factors in preventing the enzyme from inactivation were also studied. Additional experiments were performed in which the recombinant SAM synthetase was purified and immobilized in one step using immobilized metal-chelate affinity chromatography. The immobilized synthetase was found to be 40.4% of the free enzyme activity in catalyzing the synthesis of SAM from DL-Met and ATP.

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S‐Adenosylmetliionine and methylation

Cited by 810 publications

References 86 publications

AMD1 mRNA employs ribosome stalling as a mechanism for molecular memory formation

AMD1 mRNA employs ribosome stalling as a mechanism for molecular memory formation

A universal competitive fluorescence polarization activity assay for S-adenosylmethionine utilizing methyltransferases

Expression of Secreted His-Tagged S-adenosylmethionine Synthetase in the Methylotrophic Yeast Pichia pastoris and Its Characterization, One-Step Purification, and Immobilization

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