2'(3')-O-Glycyl oligoribonucleotides with sequences of the 3'-terminus of glycyl-tRNA: chemical synthesis and properties in partial reactions of protein biosynthesis

Abstract

Specific syntheses of 2'(3')-O-aminoacyl oligoribonucleotides C-C-A-Gly (12), C-C-A(AcGly) (7), U-C-C-A-Gly (17), and C-U-C-C-A-Gly (19), which are the 3'-terminal sequences of Escherichia coli Gly-tRNA (or AcGly-tRNA, respectively) are described. Compounds 12, 17, and 19 were synthesized by employing the benzotriazolyl phosphotriester approach with the following protection groups on the components: benzoyl for the heterocyclic amino groups, 2-chlorophenyl group for internucleotide phosphate protection, dimethoxytrityl and levulinoyl groups for blocking of the 5'-hydroxyl, methoxytetrahydropyranyl group for protection of the 2'-hydroxyl functions, and N-(benzyloxycarbonyl)orthoglycinate as the masked aminoacyl group simultaneously protecting the 2',3'-cis diol group of the 3'-terminal adenosine moiety. The fully protected tri-, tetra-, and pentanucleotides were obtained via 5'-extension of di- and trinucleotide blocks after prior selective removal of the 5'-O-levulinoyl group with hydrazine. The blocked derivatives 11, 16, and 18 were totally deprotected by reactions with NH4OH, H+, and H2/Pd to yield the target compounds 12, 17, and 19 in good yields. C-C-A(AcGly) (7) was synthesized according to a stepwise procedure via activation of preformed diesters with (mesitylenesulfonyl)tetrazole. C-C-A-Gly (12), U-C-C-A-Gly (17), and C-U-C-C-A-Gly (19) were all acceptor substrates in the peptidyltransferase reaction with the Ac-Phe-tRNA-70S ribosome-poly(U) system. All three models also promoted EF-Tu-70S ribosome GTP hydrolysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Levulinic acid in organic synthesis

“…The increase in the size of the hydrocarbon fragment of the primary alcohol does not impede its esterification. 48 Since primary alcohols are easily and selectively acylated under these conditions, LA is used as a protective group in the synthesis of oligonucleotides 49 and oligosaccharides. 50 ± 52 Deprotection can be achieved using sodium borohydride 53 or sulfite anion donors.…”

Site-specific incorporation of nonnatural residues during in vitro protein biosynthesis with semi-synthetic aminoacyl-tRNAs

“…Chemical synthesis of the requisite aminoacyl dinucleotide ( 9) is based upon prior work developed by Chládek and coworkers (Hagen et al, 1988;Happ et al, 1987;Scalfi-Happ et al, 1987). However, 2,-deoxycytidine is employed rather than cytidine, resulting in the synthesis of a dinucleotide RNA/DNA hybrid (Scheme I)-the details of which are reported elsewhere (Bain et al, 1991b).…”

Effect of nucleotide substitution on the peptidyltransferase activity of 2'(3')-O-(aminoacyl) oligonucleotides

“…General methods, sources of reagents, and biochemicals are the same as described in previous papers of this series (Chládek et al, 1974;Bhuta et al, 1982;Scalfi-Happ et al, 1987).…”

“…It is established that the peptidyltransferase activity increases in the order A-Gly < C-A-Gly < C-C-A-Gly, whereas activities of U-C-C-A-Gly and C-U-C-C-A-Gly are roughly similar to that of C-C-A-Gly (Bhuta et al, 1982;Scalfi-Happ et al, 1987). Similarly, binding activities to the peptidyltransferase A site are on the order of C-A-Phe < C-C-A-Phe = C-A-C-C-A-Phe (Bhuta et al, 1982).…”