Large Scale Synthesis of the Cap Part in Messenger RNA Using a New Type of Bifunctional Phosphorylating Reagent

Fukuoka, Koichiro; Suda, Fuminori; Suzuki, Ryo; Ishikawa, Masahide; Takaku, Hiroshi; Hata, Tsujiaki

doi:10.1080/15257779408012171

Cited by 17 publications

(13 citation statements)

References 13 publications

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“…Similar approach was applied previously for the synthesis of unlabeled mRNA cap analogues, where 3-fold molar excess of "cold" methyl iodide was used. [5,6] After 6 hours the reaction was quenched at 50% yield of the desired product ([7-3 H]-m 7 Gp 3 G), 20% of the double methylated by-product ([7-3 H]-m 7 Gp 3 m 7 G), and 25% of unreacted Gp 3 G, as shown by HPLC (Figure 2). Both radiolabeled products were purified using preparative RP HPLC, and their specific activity after lyophilization was determined using a scintillation counter and liquid scintillators for the aqueous samples (8.4 MBq/mg).…”

Section: Resultsmentioning

confidence: 99%

Synthesis of³H and¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Jemielity¹,

Stolarski²,

Darżynkiewicz³

2007

Nucleosides, Nucleotides and Nucleic Acids

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For deeper understanding the roles of the mRNA cap structure in cellular processes isotopically labeled dinucleotide cap analogues have been synthesized as tools for NMR and in vivo studies. Tritium or carbon C-13 labeled methyl iodide was used as a source of the isotope material. In order to minimize the number of steps during the radioisotopic synthesis the methylation with tritium labeled methyl iodide was performed with Gp(3)G as a substrate. The C-13 isotope was introduced into the cap dinucleotide by methylation of GDP with C-13 Methyl iodide, followed by coupling the product with guanosine 5'-phosphorimidazolide in DMF with zinc chloride as a catalyst.

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Section: Resultsmentioning

confidence: 99%

Synthesis of³H and¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Jemielity¹,

Stolarski²,

Darżynkiewicz³

2007

Nucleosides, Nucleotides and Nucleic Acids

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“…7-Methyl-GTP Sepharose 4B and Glutathione Sepharose 4B were purchased from Pharmacia Biotech Inc. 7-Methyl-GTP (m 7 GTP) and GTP were purchased from Sigma Chemical. m 7 GpppA was synthesized according to previously reported procedures {19, 20). Anti-eIF4E serum (rabbit) was purchased from Sawady Technology.…”

Section: Methodsmentioning

confidence: 99%

Binding Analysis of Xenopus laevis Translation Initiation Factor 4E (eIF4E) in Initiation Complex Formation

Mizumoto

Youtani

Ide

et al. 1999

Journal of Biochemistry

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A translation initiation factor, eIF4E, of Xenopus laevis was purified by affinity column chromatography after the gene expression as a full-length protein in a baculovirus-insect cell system. Interaction between X. laevis eIF4E and 4E-BP2 was analyzed by affinity column chromatography, gel permeation chromatography (GPC), and surface plasmon resonance (SPR). It was found that the interaction of eIF4E with an mRNA cap-analogue enhanced the binding activity of eIF4E with 4E-BP2. Furthermore, the SPR analysis showed that the eIF4E-cap-analogue interaction was very weak regardless of complex formation of 4E-BP2 with eIF4E; the dissociation constant of eIF4E for the cap-analogue was estimated to be 10(-2)-10(-4) M. These results suggest that the participation of another initiation factor is required for eIF4E to recognize the cap structure in vivo. The results reported in this paper support "the performed complex model" of Lee et al., in which eIF4E binds to the mRNA cap structure after the initiation factors have formed the initiation complex eIF4F.

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“…However, the reaction takes longer reaction time and results in poor yields due to poor solubility of guanosine nucleotide in the presence of organic solvent. The N7 methylation of guanosine moiety has also been achieved by the use of methyl methanesulfonate as a methylating agent under aqueous conditions (Fukuoka et al., ). We have reported an efficient synthesis of 7‐methylguanosine 5′‐ O ‐diphosphate in good yields that involves methylation of guanosine 5′‐ O ‐diphosphate using dimethyl sulfate as a methylating agent under anhydrous conditions (Kore & Parmar, ).…”

Section: Commentarymentioning

confidence: 99%

Highly Regioselective Methylation of Guanosine Nucleotides: An Efficient Synthesis of 7‐Methylguanosine Nucleotides

Senthilvelan

Shanmugasundaram

Kore

2019

CP Nucleic Acid Chemistry

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This article describes a simple, reliable, efficient, and general method for the synthesis of 7‐methylguanosine nucleotides such as 7‐methylguanosine 5′‐O‐monophosphate (m7GMP), 7‐methylguanosine 5′‐O‐diphosphate (m7GDP), 7‐methyl‐2′‐deoxyguanosine 5′‐O‐triphosphate (m72′dGTP), and 7‐methylguanosine 5′‐O‐triphosphate (m7GTP) starting from the corresponding guanosine nucleotide is described. The present protocol involves methylation reaction of guanosine nucleotide using dimethyl sulfate as a methylating agent and water as a solvent at room temperature to provide the corresponding 7‐methylguanosine nucleotide in good yields with high purity (>99.5%). It is noteworthy that the present methylation reaction proceeds smoothly under aqueous conditions that is highly regioselective to afford exclusive 7‐methylguanosine nucleotide. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Synthesis of 7‐methylguanosine nucleotides.

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Large Scale Synthesis of the Cap Part in Messenger RNA Using a New Type of Bifunctional Phosphorylating Reagent

Cited by 17 publications

References 13 publications

Synthesis of³H and¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Synthesis of³H and¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Binding Analysis of Xenopus laevis Translation Initiation Factor 4E (eIF4E) in Initiation Complex Formation

Highly Regioselective Methylation of Guanosine Nucleotides: An Efficient Synthesis of 7‐Methylguanosine Nucleotides

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Large Scale Synthesis of the Cap Part in Messenger RNA Using a New Type of Bifunctional Phosphorylating Reagent

Cited by 17 publications

References 13 publications

Synthesis of3H and13C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Synthesis of3H and13C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Binding Analysis of Xenopus laevis Translation Initiation Factor 4E (eIF4E) in Initiation Complex Formation

Highly Regioselective Methylation of Guanosine Nucleotides: An Efficient Synthesis of 7‐Methylguanosine Nucleotides

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Synthesis of³H and¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Synthesis of³H and¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies