Using S-adenosyl-l-homocysteine capture compounds to characterize S-adenosyl-l-methionine and S-adenosyl-l-homocysteine binding proteins

S -Adenosyl-L-methionine (SAM) is a sulfonium molecule with a structural hybrid of methionine and adenosine. As the second largest cofactor in the human body, its major function is to serve as methyl donor for SAM-dependent methyltransferases (MTases). The resultant transmethylation of biomolecules constitutes a significant biochemical mechanism in epigenetic regulation, cellular signaling, and metabolite degradation. Recently, numerous SAM analogs have been developed as synthetic cofactors to transfer the activated groups on MTase substrates for downstream ligation and identification. Meanwhile, new compounds built upon or derived from the SAM scaffold have been designed and tested as selective inhibitors for important MTase targets. Here, we summarized the recent development and application of SAM analogs as chemical biology tools for MTases.

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“…Compound 55 which is a fluorescein conjugate was used to measure binding affinity via fluorescence anisotropy. 139 …”

Section: Development Of Sam Analogs As Capture Compounds and Fluormentioning

confidence: 99%

SAM/SAH Analogs as Versatile Tools for SAM-Dependent Methyltransferases

2015

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“…We envisaged that the rigid triazole linker could also contribute to the ligand interaction with the enzyme. Previously, fluorescently labeled AdoMet analogs have been made with a fluorophore attached to the N6 position of the adenyl ring or the ribose hydroxyls 21-23 . However, there are no reports that describe ligands with a fluorophore tethered to the junction between the adenosyl and the 2-aminobutanoate moieties of Aza-adenosylhomocysteine.…”

Section: Resultsmentioning

confidence: 99%

Design of a fluorescent ligand targeting the S-adenosylmethionine binding site of the histone methyltransferase MLL1

Luan

Blazer

et al. 2016

Org. Biomol. Chem.

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The histone methyltransferase MLL1 has been linked to translocation-associated gene fusion in childhood leukemias and is an attractive drug target. High-throughput biochemical analysis of MLL1 methyltransferase activity requires the production of at least a trimeric complex of MLL1, RbBP5 and WDR5 to elicit robust activity. Production of trimeric and higher order MLL1 complexes in the quantities and reproducibility required for high-throughput screening presents a significant impediment to MLL1 drug discovery efforts. We present here a small molecule fluorescent ligand (FL-NAH, 6) that is able to bind to the S-adenosylmethionine (SAM) binding site of MLL1 in a manner independent of the associated complex members. We have used FL-NAH to develop a fluorescence polarization-based SAM displacement assay in a 384-well format targeting the MLL1 SET domain in the absence of associated complex members. FL-NAH competes with SAM and is displaced from the MLL1 SET domain by other SAM-binding site ligands with Kdisp values similar to the higher-order complexes, but is unaffected by the H3 peptide substrate. This assay enables screening for SAM-competitive MLL1 inhibitors without requiring the use of trimeric or higher order MLL1 complexes, significantly reducing screening time and cost.

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“…More specifically, estrogns can lower plasma homocysteine levels by formating of estrogen-homocysteine conjugates [29]. Additionally, catechol-O-methyltransferase (COMT) is responsible for both converting S-adenosyl-homocysteine to homocysteine and metabolizing catecholestrogens to methoxyestrogens and catabolize catecholamines [30,31]. Estrogens can also lower plasma homocysteine levels by competing with S-adenosyl-homocysteine for COMT.…”

Section: Discussionmentioning

confidence: 99%

Hyperhomocysteinaemia is an independent risk factor for peripheral arterial disease in a Chinese Han population

et al. 2017

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Introduction Hyperhomocysteineamia (HHcy) has long been suggested as a risk factor for atherosclerosis. However the association between HHcy and peripheral arterial disease (PAD) is still controversial. There is a lack of research on this topic in the Chinese population. This study aims to provide further results. Methods 240 PAD patients and 240 control subjects were evaluated for both serum total homocysteine levels and ankle brachial indexes (ABIs). Multivariable logistic regression models were used to estimate the association between HHcy and the risk of developing PAD. Interaction and stratified analyses were conducted according to age, sex, smoking status, drinking status, and histories of chronic disease. Results The multivariate logistic analyses revealed that the risk of PAD was significantly associated with serum homocysteine levels. The interaction analysis showed no interactive role in the association between HHcy and PAD, indicating that homocysteine was associated with PAD independent of classical vascular risk factors. Conclusion In conclusion, HHcy is an independent risk factor for PAD in the Chinese Han population. A prospective and randomized clinical trial of homocysteine lowering therapy in the Chinese population is needed to assess the causal nature of the relationship.

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Using S-adenosyl-l-homocysteine capture compounds to characterize S-adenosyl-l-methionine and S-adenosyl-l-homocysteine binding proteins

Cited by 11 publications

References 30 publications

SAM/SAH Analogs as Versatile Tools for SAM-Dependent Methyltransferases

SAM/SAH Analogs as Versatile Tools for SAM-Dependent Methyltransferases

Design of a fluorescent ligand targeting the S-adenosylmethionine binding site of the histone methyltransferase MLL1

Hyperhomocysteinaemia is an independent risk factor for peripheral arterial disease in a Chinese Han population

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