Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase

Law, Brian J. C.; Struck, Anna‐Winona; Bennett, Matthew R.; Wilkinson, Barrie; Micklefield, Jason

doi:10.1039/c5sc00164a

Cited by 52 publications

(64 citation statements)

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“…Selective methylation processes are of great interest for syntheses of bioactive substances . Methylation of drug molecules has often been described as very important for their efficiency and bioactivity .…”

Section: Methodsmentioning

confidence: 99%

“…The results of this study are important milestones on the pathway to catalytic SAM-dependentalkylation processes.Selectivem ethylation processes are of great interest for syntheses of bioactive substances. [1][2][3][4] Methylation of drug molecules has often been describeda sv ery important for their efficiency and bioactivity. [5,6] In nature,c hemo-, regio-and stereoselectivem ethylations are mainly accomplished through S-adenosylmethionine (SAM)-dependentm ethyltransferases (MTs) that provide exceptional access to C-, N-, O-and S-methylated compounds as wella st oh alomethanes.…”

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confidence: 99%

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Regiocomplementary O‐Methylation of Catechols by Using Three‐Enzyme Cascades

et al. 2015

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S-Adenosylmethionine (SAM)-dependent enzymes have great potential for selective alkylation processes. In this study we investigated the regiocomplementary O-methylation of catechols. Enzymatic methylation is often hampered by the need for a stoichiometric supply of SAM and the inhibitory effect of the SAM-derived byproduct on most methyltransferases. To counteract these issues we set up an enzyme cascade. Firstly, SAM was generated from l-methionine and ATP by use of an archaeal methionine adenosyltransferase. Secondly, 4-O-methylation of the substrates dopamine and dihydrocaffeic acid was achieved by use of SafC from the saframycin biosynthesis pathway in 40-70 % yield and high selectivity. The regiocomplementary 3-O-methylation was catalysed by catechol O-methyltransferase from rat. Thirdly, the beneficial influence of a nucleosidase on the overall conversion was demonstrated. The results of this study are important milestones on the pathway to catalytic SAM-dependent alkylation processes.

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Section: Methodsmentioning

confidence: 99%

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confidence: 99%

Regiocomplementary O‐Methylation of Catechols by Using Three‐Enzyme Cascades

et al. 2015

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“…13–15,27,28 While there have been great advances in methylation-dependent bioinformatics and disease-associated biomarkers, 29–31 the study of intracellular MT spatial/temporal resolution, specificity, and/or function remains a challenge. 9,14,15,25–28,32–36 Toward this end, the pioneering demonstration of AdoMet analogs, bearing alternative alkyl donor substituents, as cosubstrates for DNA 37 or natural product (NP) 38 MTs has inspired new tools and strategies to study NP, 39–42 protein, 43–51 and nucleic acid 6,52–57 methylation where the recent development of enzyme-based strategies for the synthesis of differentially alkylated AdoMet analogs has simplified access to these unique cosubstrates. 42,49,50,57–59 However, the stability of AdoMet or its corresponding differentially alkylated analogs under physiological conditions limits their utility as reagents or therapeutics by virtue of two fundamental degradative processes: intramolecular cyclization to homoserine lactone and 5′-deoxy-5′-(alkylthio)adenosine (Figure 1, pathway a) and depurination (Figure 1, pathway b).…”

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Functional AdoMet Isosteres Resistant to Classical AdoMet Degradation Pathways

et al. 2016

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S-adenosyl-l-methionine (AdoMet) is an essential enzyme cosubstrate in fundamental biology with an expanding range of biocatalytic and therapeutic applications. We report the design, synthesis, and evaluation of stable, functional AdoMet isosteres that are resistant to the primary contributors to AdoMet degradation (depurination, intramolecular cyclization, and sulfonium epimerization). Corresponding biochemical and structural studies demonstrate the AdoMet surrogates to serve as competent enzyme cosubstrates and to bind a prototypical class I model methyltransferase (DnrK) in a manner nearly identical to AdoMet. Given this conservation in function and molecular recognition, the isosteres presented are anticipated to serve as useful surrogates in other AdoMet-dependent processes and may also be resistant to, and/or potentially even inhibit, other therapeutically relevant AdoMet-dependent metabolic transformations (such as the validated drug target AdoMet decarboxylase). This work also highlights the ability of the prototypical class I model methyltransferase DnrK to accept non-native surrogate acceptors as an enabling feature of a new high-throughput methyltransferase assay.

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“…[20,21] One limitation of this process is the need to prepare these cofactors by chemical synthesis,w hich is laborious,l ow yielding,a nd produces both epimers at the sulfur center. [15,29,30] One example of this one-pot process is the generation of cofactor analogues in situ from either ClDAo rA TP and (m)ethionine, [15,[29][30][31][32][33] followed by C-(m)ethyl transfer catalyzed by aM Tase (Figure 1b). [27,28] Am ore step-and atom-efficient strategy is to couple cofactor formation with C-alkyl transfer.…”

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confidence: 99%

S‐Adenosyl Methionine Cofactor Modifications Enhance the Biocatalytic Repertoire of Small Molecule C‐Alkylation

et al. 2019

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Atandem enzymatic strategy to enhance the scope of C-alkylation of small molecules via the in situ formation of Sadenosyl methionine (SAM) cofactor analogues is described. As olvent-exposed channel present in the SAM-forming enzyme SalL tolerates 5'-chloro-5'-deoxyadenosine (ClDA) analogues modified at the 2-position of the adenine nucleobase.C oupling SalL-catalyzed cofactor production with C-(m)ethyl transfer to coumarin substrates catalyzedb yt he methyltransferase (MTase) NovOf orms C-(m)ethylated coumarins in superior yield and greater substrate scope relative to that obtained using cofactors lacking nucleobase modifications.E stablishing the molecular determinants that influence C-alkylation provides the basis to develop al ate-stage enzymatic platform for the preparation of high value small molecules.

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Site-specific bioalkylation of rapamycin by the RapM 16-O-methyltransferase

Abstract: Characterisation of a rapamycin O-methyltransferase (RapM) and its utilisation in coupled reactions, with an improved variant of the human methionine adenosyl transferase (hMAT2A), results in new regioselectively alkylated rapamycin derivatives.

Cited by 52 publications

References 50 publications

Regiocomplementary O‐Methylation of Catechols by Using Three‐Enzyme Cascades

Regiocomplementary O‐Methylation of Catechols by Using Three‐Enzyme Cascades

Functional AdoMet Isosteres Resistant to Classical AdoMet Degradation Pathways

S‐Adenosyl Methionine Cofactor Modifications Enhance the Biocatalytic Repertoire of Small Molecule C‐Alkylation

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