Synthesis of ³H and ¹³C Labeled Mrna Cap Dinucleotides—Useful Tools for Nmr, Biochemical, and Biological Studies

Abstract

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 Gp₃G 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.

Keywords:

• 5′ mRNA Cap
• tritium
• carbon 13
• isotope labeling

This work was supported by grant from HHMI (No. 55005604) and grants from Polish Ministry of Science and Higher Education (No. 3 P04A 021 25 and No. 2 P04A 006 28).