Quantitative Labeling of Long Plasmid DNA with Nanometer Precision

Pljevaljčić, Goran; Schmidt, Falk; Scheidig, Axel J.; Weinhold, Elmar G.

doi:10.1002/cbic.200700294

Cited by 27 publications

(24 citation statements)

References 18 publications

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“…Synthesis of the SAH derivative with an amino linker at C8 started from 8-bromo-2',3'-O-isopropylideneadenosine (9). Initially, we followed the synthetic route for the N6-modified SAH derivative 8.…”

Section: Resultsmentioning

confidence: 99%

“…[7] However, based on our experience with aziridinebased SAM analogues for labeling DNA using MTases, [8] X-ray structures of MTases in complex with these cofactor analogues [9] and analysis of PDB data, [10] the exocyclic N6 position or C8 position of the adenine heterocycle (Scheme 1) are much less recognized by MTases and more suitable for linker introduction and subsequent attachment of CC scaffolds. To target these positions we implemented new synthetic routes to adenine-modified SAH derivatives.…”

Section: Introductionmentioning

confidence: 99%

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Synthesis of S‐Adenosyl‐L‐homocysteine Capture Compounds for Selective Photoinduced Isolation of Methyltransferases

Dalhoff¹,

Hüben

Lenz³

et al. 2010

ChemBioChem

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Understanding the interplay of different cellular proteins and their substrates is of major interest in the postgenomic era. For this purpose, selective isolation and identification of proteins from complex biological samples is necessary and targeted isolation of enzyme families is a challenging task. Over the last years, methods like activity-based protein profiling (ABPP) and capture compound mass spectrometry (CCMS) have been developed to reduce the complexity of the proteome by means of protein function in contrast to standard approaches, which utilize differences in physical properties for protein separation. To isolate and identify the subproteome consisting of S-adenosyl-L-methionine (SAM or AdoMet)-dependent methyltransferases (methylome), we developed and synthesized trifunctional capture compounds containing the chemically stable cofactor product S-adenosyl-L-homocysteine (SAH or AdoHcy) as selectivity function. SAH analogues with amino linkers at the N6 or C8 positions were synthesized and attached to scaffolds containing different photocrosslinking groups for covalent protein modification and biotin for affinity isolation. The utility of these SAH capture compounds for selective photoinduced protein isolation is demonstrated for various methyltransferases (MTases) acting on DNA, RNA and proteins as well as with Escherichia coli cell lysate. In addition, they can be used to determine dissociation constants for MTase-cofactor complexes.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Synthesis of S‐Adenosyl‐L‐homocysteine Capture Compounds for Selective Photoinduced Isolation of Methyltransferases

Dalhoff¹,

Hüben

Lenz³

et al. 2010

ChemBioChem

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“…[44] This group employed methyltransferases (MTases), which catalyze the transfer of an activated methyl group from the ubiquitous cofactor Sadenosyl-l-methinone (AdoMet or SAM) to adenine or cytosine residues on DNA within specific DNA sequences between two and eight base pairs long. [45] Similar to Weinhold and co-workers, [44] Weller and Rajski prepared the novel cofactor 26, which is accepted by the methyltransferase M.TaqI instead of natural SAM (Scheme 8). With the novel cofactor, however, not a methyl group but a propargylmodified adenine moiety was transferred to the N6 nitrogen of adenine in the palindromic sequence 5'-AGCT-3'.…”

Section: Dna Click Chemistrymentioning

confidence: 96%

Postsynthetic DNA Modification through the Copper‐Catalyzed Azide–Alkyne Cycloaddition Reaction

et al. 2008

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The attachment of labels onto DNA is of utmost importance in many areas of biomedical research and is valuable in the construction of DNA-based functional nanomaterials. The copper(I)-catalyzed Huisgen cycloaddition of azides and alkynes (CuAAC) has recently been added to the repertoire of DNA labeling methods, thus allowing the virtually unlimited functionalization of both small synthetic oligonucleotides and large gene fragments with unprecedented efficiency. The CuAAC reaction yields the labeled polynucleotides in very high purity after a simple precipitation step. The reviewed technology is currently changing the way in which functionalized DNA strands are generated cost-efficiently in high quality for their application in molecular diagnostics systems and nanotechnological research.

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“…[44] Weinhold et al verwendeten Methyltransferasen (MTasen), die den Transfer aktivierter Methylgruppen vom ubiquitären Cofaktor S-Adenosyl-l-methionin (AdoMet oder SAM) auf Adenin-oder Cytosinreste in der DNA in- nerhalb spezifischer Sequenzen von zwei bis acht Basenpaaren Länge katalysieren. [45] ¾hnlich wie Weinhold et al [44] stellten …”

Section: Mehrfache Sequenzielle Markierung Von Dnaunclassified

Postsynthetische DNA‐Modifizierung mithilfe der kupferkatalysierten Azid‐Alkin‐Cycloaddition

et al. 2008

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Die Markierung von DNA ist von größter Bedeutung für die Biomedizin und bildet die Basis für die Konstruktion DNA‐basierter funktionaler Nanomaterialien. Seit kurzem zählt die Kupfer(I)‐katalysierte Huisgen‐Cycloaddition von Aziden mit Alkinen (CuAAC) zum Repertoire der DNA‐Markierungsmethoden. Diese Methode ermöglicht die Modifizierung von DNA mit beispielloser Effizienz. Nicht nur kleine Oligonucleotide, sondern auch ganze Genfragmente können so hochgradig chemisch modifiziert werden. Die CuAAC‐Reaktion liefert die markierten Polynucleotide, meist nach einfacher Ausfällung, in ausgezeichneter Reinheit und ist auf dem besten Wege, die Synthese funktionalisierter DNA‐Stränge zu revolutionieren. Die Methode wird von entscheidender Bedeutung für die Erzeugung neuer diagnostischer Systeme und für die nanotechnologische Forschung sein.

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Quantitative Labeling of Long Plasmid DNA with Nanometer Precision

Cited by 27 publications

References 18 publications

Synthesis of S‐Adenosyl‐L‐homocysteine Capture Compounds for Selective Photoinduced Isolation of Methyltransferases

Synthesis of S‐Adenosyl‐L‐homocysteine Capture Compounds for Selective Photoinduced Isolation of Methyltransferases

Postsynthetic DNA Modification through the Copper‐Catalyzed Azide–Alkyne Cycloaddition Reaction

Postsynthetische DNA‐Modifizierung mithilfe der kupferkatalysierten Azid‐Alkin‐Cycloaddition

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