Assignment of the Absolute Configuration of P-Chiral 5′Mrna Cap Analogues Containing Phosphorothioate Moiety

Abstract

Enzymatic cleavage of the P-chiral diastereoisomers of the 5′ mRNA cap analogue bearing phosphorothioate moiety in alfa position of 5′,5′-triphosphate bridge (m⁷Gppp _S G D1 and D2) was performed by human Decapping Scavenger (DcpS) enzyme. Analysis of the degradation products allowed to estimate the absolute configuration at the asymmetric phosphorus atoms in examined compounds via correlation with the R _P and S _P diastereoisomers of guanosine 5′-O-(1-thiodiphosphate) (GDPαS).

Keywords:

• 5′ mRNA Cap
• phosphorothioate
• P-chirality
• absolute configuration
• enzymatic hydrolysis

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 3 T09A 153 29). The authors thank to Dr. R. E. Davis (The University of Colorado Heath Sciences Center, Aurora, Colorado, USA) for his generous gift of the hDcpS palsmid.