Nucleoside-modified AdoMet analogues for differential methyltransferase targeting

Cornelissen, Nicolas V.; Michailidou, Freideriki; Muttach, Fabian; Rau, Kristina; Rentmeister, Andrea

doi:10.1039/c9cc07807j

Cited by 27 publications

(36 citation statements)

References 41 publications

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“…[20] Efforts to circumvent the stability issues and the need to prepare SAM analogues separately have focused on developing tandem enzymatic reactions, where the cofactor is synthesized in situ, followed by MTase-mediated methylation. [21][22][23][24][25][26][27] Another alternative is to develop shelf-stable SAM analogues (e. g., 7-9), which are accepted by MTases as substrates but could also be stored prior to use ( Figure 2B). [20] The second challenge in establishing a biocatalytic methylation platform arises from the inherent need to use stoichiometric amounts of the cofactor (or analogue), which invariably produces an equal amount of the S-adenosylhomocysteine (SAH) by-product.…”

Section: Current Challenges Of Biocatalytic Alkylationmentioning

confidence: 99%

“…[35] 4. Tandem enzymatic alkylation by using A) MAT, [9,[20][21]25,[37][38][39] B) an HMT-mediated SAM regeneration process, [40] and C) SalL to generate SAM analogues in situ.…”

Section: Development Of Sam Cofactor Regeneration Systemsmentioning

confidence: 99%

“…A pertinent exemplar of this promiscuity is DNA/RNA MTases which accept analogues of SAM and can be used as a tagging tool for biotechnologybased applications. [19,[25][26][41][42][43] Cofactor promiscuity is also a hallmark of a number of small molecule C-MTases such as NovO and CouO, which accept a variety of S-alkylated SAM analogues prepared separately by chemical synthesis. [44] A distinct limitation of preparing SAM analogues by chemical synthesis is the formation of R/S epimers at the S centre.…”

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

“…A solvent accessible active site of MAT enables this enzyme to tolerate analogues of methionine and (seleno)methionine substrates bearing longer alkyl chains and functional handles on S/Se-centres producing a suite of analogues such as 7-9 ( Figure 2). [21,25,39,[47][48][49] The in situ synthesis of these cofactor analogues can then be used as substrates to transfer functional handles catalyzed by a variety of O- (14 a-d), C-(14 e) and N-(14 g) MTases (Figure 4a). [9,[20][21]25,[37][38][39] In addition, Seebeck et al expanded their two-enzyme SAM regeneration strategy (Figure 3B) [35] to stereoselectively prepare L-or D-β-alkylated αamino acids amino acids such as 14 f ( Figure 4B), [40] which are challenging to prepare by conventional chemical synthesis.…”

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

“…The substrate promiscuity of MAT enzymes in this respect are particularly attractive for SAM analogue synthesis which have been optimized by rational design. [21,25,[48][49] Since ATP is present in cells in millimolar concentrations, there exists considerable potential to engineer cascade checkpoints by directed evolution. This could lead to developing late-stage alkylation platforms which function in live cells or cell lysates.…”

Section: Towards the Development Of An Orthogonal Biocatalytic Alkylamentioning

confidence: 99%

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Biocatalytic Alkylation Cascades: Recent Advances and Future Opportunities for Late‐Stage Functionalization

2020

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This Concept article describes the latest developments in the emerging area of late-stage biocatalytic alkylation. Central to these developments is the ability to efficiently prepare Sadenosyl methionine (SAM) cofactor analogues and couple this with enzymatic alkyl transfer. Recent developments in the enzymatic synthesis of SAM cofactor analogues are summarized first, followed by their application as alkyl transfer agents catalyzed by methyltransferases (MTases). Second, innovative methods to regenerate SAM cofactors by enzymatic cascades is reported. Finally, future opportunities towards establishing a generalized platform for late-stage alkylation are described.

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Section: Current Challenges Of Biocatalytic Alkylationmentioning

confidence: 99%

“…[35] 4. Tandem enzymatic alkylation by using A) MAT, [9,[20][21]25,[37][38][39] B) an HMT-mediated SAM regeneration process, [40] and C) SalL to generate SAM analogues in situ.…”

Section: Development Of Sam Cofactor Regeneration Systemsmentioning

confidence: 99%

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

Section: Enhancing the Scope Of Biocatalytic Alkylation Using Modifiementioning

confidence: 99%

Section: Towards the Development Of An Orthogonal Biocatalytic Alkylamentioning

confidence: 99%

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Biocatalytic Alkylation Cascades: Recent Advances and Future Opportunities for Late‐Stage Functionalization

2020

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From Stoichiometric Reagents to Catalytic Partners: Selenonium Salts as Alkylating Agents for Nucleophilic Displacement Reactions in Water

et al. 2021

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The ability of chalcogenium salts to transfer an electrophilic moiety to a given nucleophile is well known. However, up to date, these reagents have been used in stoichiometric quantities, producing a substantial amount of waste as byproducts of the reaction. In this report, we disclose further investigation of selenonium salts as Sadenosyl-L-methionine (SAM) surrogates for the alkylation of nucleophiles in aqueous solutions. Most importantly, we were able to convert the stoichiometric process to a catalytic system employing as little as 10 mol % of selenides to accelerate the reaction between benzyl bromide and other alkylating agents with sodium cyanide in water. Probe experiments including 77 Se NMR and HRMS of the reaction mixture have unequivocally shown the presence of the selenonium salt in the reaction mixture.

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Maßgeschneiderte SAM‐Synthetasen zur enzymatischen Herstellung von AdoMet‐Analoga mit Photoschutzgruppen und zur reversiblen DNA‐Modifizierung in Kaskadenreaktionen

et al. 2020

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Die Methylierung und Demethylierung von DNA, RNA und Proteinen stellt einen wichtigen Regulationsmechanismus dar. Um die Funktionen dieser Modifikationen aufzuklären, wären Werkzeuge nützlich, die die sequenzspezifische Blockierung und zeitlich präzise Freisetzung der entsprechenden Stellen ermçglichen. S-Adenosyl-l-Methionin (Ado-Met)-Analoga erweisen sich in Kombination mit Methyltransferasen (MTasen) als nützlich bei der Kartierung oder Blockierung und Freisetzung von MTase-Zielsequenzen. Allerdings war ihre enzymatische Herstellung bisher auf aliphatische Gruppen am Schwefelatom beschränkt. Wir konnten eine SAM-Synthetase aus Cryptosporidium hominis (PS-ChMAT) für die effiziente Herstellung von AdoMet-Analoga mit Photoschutzgruppen entwickeln, die von keiner der bisher bekannten WT-MATs akzeptiert werden. Die Kristallstruktur von PS-ChMAT bei 1.87 zeigt, wie das Photoschutzgruppen tragende AdoMet-Analogon in der Enzymtasche vorliegt und bahnte den Weg für die Entwicklung einer thermostabilen MAT aus Methanocaldococcus jannaschii. Die PS-MATs sind mit DNA-und RNA-MTasen kompatibel und ermçglichen die sequenzspezifische Modifizierung einer Plasmid-DNA ("Schreiben") sowie die lichtgesteuerte Entfernung ("Lçschen"). Einleitung Die Methylierung von DNA, RNA und Histonen ist oft reversibel und stellt einen Regulationsmechanismus dar, der direkte Auswirkungen auf grundlegende biologische Prozesse und menschliche Krankheiten hat. [1] Diese epigenetische Markierung wird durch Methyltransferasen (MTasen) eingeführt, die typischerweise S-Adenosyl-l-Methionin (SAM oder AdoMet) als Methyldonor verwenden. [2] In der DNA führt m 5 C (5-Methylcytosin) zur Inaktivierung der Transkriptions-Start-Sequenzen, während die oxidative Entfer-nung die Genexpression wiederherstellt. Kürzlich wurde gezeigt, dass m 6 A (N 6-Methyladenosin) in der DNA an der Aktivierung und Hemmung der Transkription beteiligt ist. [2d, 3] Die Fähigkeit, solche Methyltransferase-Zielsequenzen zu blockieren und sie zu einem definierten Zeitpunkt mit einem orthogonalen Auslçser freizusetzen, würde eingehende Studien ermçglichen, die erforderlich sind, um die biologische Funktion im Detail zu verstehen. [1d] Photoschutzgruppen sind wirkungsvolle Werkzeuge zur Untersuchung biomolekularer Wechselwirkungen und Funktionen. [4] Sie wurden erfolgreich für DNA, RNA und Proteine in vitro, in Zellen und in vivo eingesetzt. [4a,b] Ihre Entfernung durch Licht rekonstituiert das native Biomolekül und lässt sich mit ausgezeichneter räumlicher und zeitlicher Präzision steuern. Die 2-Nitrobenzyl-Gruppe (ONB) und ihre Derivate sind aufgrund ihrer hohen Stabilität und leichten Darstellbarkeit weit verbreitet. [5] Wir und andere konnten kürzlich zeigen, dass benzylische [6] und Photoschutz-Gruppen (PS) von AdoMet-Analoga [5b, 7] mittels sterisch nicht eingeschränkter MTasen übertragen werden kçnnen und somit zur Blockierung der Zielsequenzen dieser Enzyme geeignet sind. Die Aktivität einer MTase zur Installation einer PS-Gruppe ("Schreiben") kann also mit Licht zur Entfernung ...

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Nucleoside-modified AdoMet analogues for differential methyltransferase targeting

Abstract: Methyltransferases modify a wide range of biomolecules using S-adenosyl-l-methionine (AdoMet) as cosubstrate. Enzymatic generation of nucleoside-modified AdoMet analogues and conversion by different methyltransferases is shown.

Cited by 27 publications

References 41 publications

Biocatalytic Alkylation Cascades: Recent Advances and Future Opportunities for Late‐Stage Functionalization

Biocatalytic Alkylation Cascades: Recent Advances and Future Opportunities for Late‐Stage Functionalization

From Stoichiometric Reagents to Catalytic Partners: Selenonium Salts as Alkylating Agents for Nucleophilic Displacement Reactions in Water

Maßgeschneiderte SAM‐Synthetasen zur enzymatischen Herstellung von AdoMet‐Analoga mit Photoschutzgruppen und zur reversiblen DNA‐Modifizierung in Kaskadenreaktionen

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