Synthesis of O6-p-nitrophenylethyl guanosine and 2′-deoxyguanosine derivatives

Trichtinger, Thomas; Charubala, Ramamurthy; Pfleiderer, Wolfgang

doi:10.1016/s0040-4039(00)81505-1

Cited by 51 publications

(26 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…After washing steps, the monomethoxytrityl group at the 5' position was removed with 5% CC13COOH. The (20,21). Furthermore, the pnitrophenylethyl group is removed concomitantly with the alkylsilyl groups by using the Bu4NF/tetrahydrofuran reagent.…”

Section: Methodsmentioning

confidence: 99%

Total chemical synthesis of a 77-nucleotide-long RNA sequence having methionine-acceptance activity.

Ogilvie

Usman

Nicoghosian

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

153

View full text Add to dashboard Cite

Chemical synthesis is described of a 77-nucleotide-long RNA molecule that has the sequence of an Escherichia coli Ado47-containing tRNAP41t species in which the modified nucleosides have been substituted by their unmodified parent nucleosides. The sequence was assembled on a solid-phase, controlled-pore glass support in a stepwise manner with an automated DNA synthesizer. The ribonucleotide building blocks used were fully protected 5'-monomethoxytrityl-2'-silyl-3'-N,N-diisopropylaminophosphoramidites. p-Nitrophenylethyl groups were used to protect the o' of guanine residues. The fully deprotected tRNA analogue was characterized by polyacrylamide gel electrophoresis (sizing), terminal nucleotide analysis, sequencing, and total enzyme degradation, all of which indicated that the sequence was correct and contained only 3-5 linkages. The 77-mer was then assayed for amino acid acceptor activity by using E. coli methionyl-tRNA synthetase. The results indicated that the synthetic product, lacking modified bases, is a substrate for the enzyme and has an amino acid acceptance 11% of that of the major native species, tRNA e't containing 7-methylguanosine at position 47.Ribonucleic acids (RNAs) play a wide range of important roles in living cells. These include the traditional roles in transcription and translation (mRNA, tRNA, and rRNA) of genetic information. Excitement in RNA chemistry has grown with the discovery of RNA sequences with catalytic activity (1), the formation of "lariats" during the processing of pre-RNA (2), and the development of recombinant RNA technology (3, 4). RNA sequences of the tRNA type are required in the first step of chlorophyll biosynthesis (5). tRNAs are particularly attractive as synthetic targets because their length, 70-85 nucleotides, is sufficient to demonstrate a successful chemical procedure. In addition, they have specific biological functions that can be demonstrated in the synthetic product. They are also interesting molecules in their own right, having been implicated in gene expression (6), in propagation of tumor viruses (7), and in cellular differentiation and development in cancer induction (8-10). The chemical synthesis of RNA molecules allows complete flexibility in the choice of sequence and the ability to introduce modifications at specific sites. Thus, the ability to chemically synthesize molecules of this size should permit a rapid increase in the rate of understanding of the full biological roles of RNA molecules paralleling that which occurred earlier when the chemical synthesis of DNA oligomers became routine.Our laboratory has had as a major objective the development of methods for the chemical synthesis ofRNA sequence (11,12). These methods have now progressed to the point where the synthesis of 70-to 80-unit molecules is a practical possibility. To demonstrate the chemical method, a sequence was chosen that was identical to that of an Escherichia coli Ado-47 tRNAP et species (i.e., containing adenosine instead of 7-methylguanosine at position 47), except th...

show abstract

Section: Methodsmentioning

confidence: 99%

Total chemical synthesis of a 77-nucleotide-long RNA sequence having methionine-acceptance activity.

Ogilvie

Usman

Nicoghosian

et al. 1988

Proc. Natl. Acad. Sci. U.S.A.

153

View full text Add to dashboard Cite

show abstract

“…Introduction of the O 6 -NPE group was accomplished under Mitsunobu conditions. [28] Although optimized over the years in our laboratory, we experienced unsatisfactory yields for this transformation. Additionally, extensive purification protocols (to remove triphenylphosphine oxide) were required, rendering such a pathway time consuming and not very attractive.…”

Section: Resultsmentioning

confidence: 99%

An Unconventional Acid‐Labile Nucleobase Protection Concept for Guanosine Phosphoramidites in RNA Solid‐Phase Synthesis

Jud

Micura

2017

Chemistry A European J

View full text Add to dashboard Cite

We present an innovative O 6 -tert-butyl/N 2 -tert-butyloxycarbonyl protection concept for guanosine (G) phosphoramidites. This concept is advantageous for 2′-modified G building blocks because of very efficient synthetic access when compared with existing routes that usually employ O 6 -(4-nitrophenyl)ethyl/N 2 -acyl protection or that start from 2-aminoadenosine involving enzymatic transformation into guanosine later on in the synthetic path. The new phosphoramidites are fully compatible with 2′-O-tBDMS or TOM phosphoramidites in standard RNA solid-phase synthesis and deprotection, and provide excellent quality of tailored RNAs for the growing range of applications in RNA biophysics, biochemistry, and biology.

show abstract

“…The latter reaction which leads to a modification of the base is most pronounced with guanosine (converted into 2,6-diaminopurine), and the different tested deblocking conditions gave substantial amounts of base modification. [36] To avoid this problem, O 6 -protection of guanosine [39,40] would be recommended. If O 6 -protection is employed together with O 4 -protection of uridines (or thymidines), diisopropyl chlorophosphate or OCP could perhaps be advantageous, since condensations are more rapid with these reagents than with DMOCP.…”

Section: Discussionmentioning

confidence: 99%

Investigation on Condensing Agents for Phosphinate Ester Formation with Nucleoside 5′‐Hydroxyl Functions

Winqvist

Strömberg

2008

Eur J Org Chem

View full text Add to dashboard Cite

Condensation of a uridine 3′‐deoxy‐3′‐C‐methylenephosphinate with thymidine and guanosine derivatives to form methylenephosphinate esters was investigated. A number of different condensing agents were compared, and these include pivaloyl chloride, triisopropylbenzenesulfonyl chloride (TPS‐Cl), phosphonium and uronium derivatives, numerous chlorophosphates and bis(2‐oxo‐3‐oxazolidinyl)phosphinic chloride (OXP). The phosphonium derivatives gave slow condensations or oxidative side reactions (hydroxybenzotriazole derivatives) during preactivation of the methylenephosphinate. Pivaloyl chloride gave long coupling times, and competing 5′‐O‐pivaloylation was detected. TPS‐Cl gave rapid condensation but also rapid oxidation of the product. Most chlorophosphates gave competing 5′‐O‐phosphorylation of the nucleoside component, as well as base phosphorylation. However, 2‐chloro‐5,5‐dimethyl‐2‐oxo‐1,3,2‐dioxaphosphorinane (DMOCP) gave a rather efficient formation of dinucleoside methylenephosphinates at a decent rate. However, O6‐protection of guanines could become necessary with this reagent, since upon extended reaction time traces of O6‐phosphorylation were detected even with a low concentration (60 mM) of DMOCP (2 equiv. to phosphinate). Bis(2‐oxo‐3‐oxazolidinyl)phosphinic chloride (OXP) can, unlike DMOCP, be used in nearly equimolar amounts to phosphinate. Under such conditions OXP gives virtually quantitative condensation at a rate comparable to that of 2 equiv. of DMOCP and with no side reactions detected. We could also not detect any decomposition of OXP‐preactivated phosphinate. Nucleophilic catalysts, more powerful than pyridine (N‐methylimidazole, iodide and 4‐methoxypyridine), accelerated the reactions with OXP, but preactivation in the absence of the 5′‐OH component led to decomposition of the activated phosphinate.(© Wiley‐VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2008)

show abstract

Synthesis of O6-p-nitrophenylethyl guanosine and 2′-deoxyguanosine derivatives

Cited by 51 publications

References 46 publications

Total chemical synthesis of a 77-nucleotide-long RNA sequence having methionine-acceptance activity.

Total chemical synthesis of a 77-nucleotide-long RNA sequence having methionine-acceptance activity.

An Unconventional Acid‐Labile Nucleobase Protection Concept for Guanosine Phosphoramidites in RNA Solid‐Phase Synthesis

Investigation on Condensing Agents for Phosphinate Ester Formation with Nucleoside 5′‐Hydroxyl Functions

Contact Info

Product

Resources

About