Synthesis and biological evaluation of non-isomerizable analogues of Ala-tRNAAla

Mellal, Dénia; Fonvielle, Matthieu; Santarem, Marco; Chemama, Maryline; Schneider, Yoann; Iannazzo, Laura; Braud, Emmanuelle; Arthur, Michel; Ethève-Quelquejeu, Mélanie

doi:10.1039/c3ob41206g

Cited by 5 publications

(2 citation statements)

References 46 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Unnatural amino acids charged on tRNAs have found wide application as substrates for studying the mechanism of protein synthesis, investigating protein function by mutagenesis and introduction of biophysical probes, and synthesizing peptidomimetics for drug discovery. − In some applications, it is advantageous to alter the 3′-terminal ribose. − However, the most flexible method in terms of the amino acid portion of unnatural aminoacyl-tRNAs and potential applications is a semisynthetic methodology based on chemical aminoacylation of dinucleotide pdCpA, and its subsequent enzymatic ligation to a truncated tRNA fragment, tRNA minus CA …”

Section: Introductionmentioning

confidence: 99%

Facile Synthesis of N-Acyl-aminoacyl-pCpA for Preparation of Mischarged Fully Ribo tRNA

2014

View full text Add to dashboard Cite

Chemical synthesis of N-acyl-aminoacyl-pdCpA and its ligation to tRNA(minus CA) is widely used for the preparation of unnatural aminoacyl-tRNA substrates for ribosomal translation. However, the presence of the unnatural deoxyribose can decrease incorporation yield in translation and there is no straightforward method for chemical synthesis of the natural ribo version. Here, we show that pCpA is surprisingly stable to treatment with strong organic bases provided that anhydrous conditions are used. This allowed development of a facile method for chemical aminoacylation of pCpA. Preparative synthesis of pCpA was also simplified by using t-butyl-dithiomethyl protecting group methodology, and a more reliable pCpA postpurification treatment method was developed. Such aminoacyl-pCpA analogues ligated to tRNA(minus CA) transcripts are highly active in a purified translation system, demonstrating utility of our synthetic method.

show abstract

Section: Introductionmentioning

confidence: 99%

Facile Synthesis of N-Acyl-aminoacyl-pCpA for Preparation of Mischarged Fully Ribo tRNA

2014

View full text Add to dashboard Cite

show abstract

“…The synthesis started by addition of commercially available diethylsquarate to the 2′‐deoxy‐2′‐amino‐adenosine 1 in basic conditions. The reaction provided 2′‐squaramate adenosine 2 in 80 % yield.…”

Section: Figurementioning

confidence: 99%

Electrophilic RNA for Peptidyl‐RNA Synthesis and Site‐Specific Cross‐Linking with tRNA‐Binding Enzymes

et al. 2016

Self Cite

View full text Add to dashboard Cite

RNAf unctionalization is challenging due to the instability of RNAand the limited range of available enzymatic reactions.W ed eveloped as trategy based on solid phase synthesis and post-functionalization to introduce an electrophilic site at the 3' end of tRNAanalogues.The squarate diester used as an electrophile enabled sequential amidation and provided asymmetric squaramides with high selectivity.T he squaramate-RNAs specifically reacted with the lysine of UDP-MurNAc-pentapeptide,apeptidoglycan precursor used by the aminoacyl-transferase FemX Wv for synthesis of the bacterial cell wall. The peptidyl-RNAo btained with squaramate-RNA and unprotected UDP-MurNAc-pentapeptide efficiently inhibited FemX Wv .T he squaramate unit also promoted specific cross-linking of RNAtothe catalytic LysofFemX Wv but not to related transferases recognizing different aminoacyl-tRNAs. Thus,s quaramate-RNAs provide specificity for cross-linking with defined groups in complex biomolecules due to its unique reactivity.

show abstract

Electrophilic RNA for Peptidyl‐RNA Synthesis and Site‐Specific Cross‐Linking with tRNA‐Binding Enzymes

et al. 2016

View full text Add to dashboard Cite

RNA functionalization is challenging due to the instability of RNA and the limited range of available enzymatic reactions. We developed a strategy based on solid phase synthesis and post-functionalization to introduce an electrophilic site at the 3' end of tRNA analogues. The squarate diester used as an electrophile enabled sequential amidation and provided asymmetric squaramides with high selectivity. The squaramate-RNAs specifically reacted with the lysine of UDP-MurNAc-pentapeptide, a peptidoglycan precursor used by the aminoacyl-transferase FemX for synthesis of the bacterial cell wall. The peptidyl-RNA obtained with squaramate-RNA and unprotected UDP-MurNAc-pentapeptide efficiently inhibited FemX . The squaramate unit also promoted specific cross-linking of RNA to the catalytic Lys of FemX but not to related transferases recognizing different aminoacyl-tRNAs. Thus, squaramate-RNAs provide specificity for cross-linking with defined groups in complex biomolecules due to its unique reactivity.

show abstract

Synthesis and biological evaluation of non-isomerizable analogues of Ala-tRNAAla

Cited by 5 publications

References 46 publications

Facile Synthesis of N-Acyl-aminoacyl-pCpA for Preparation of Mischarged Fully Ribo tRNA

Facile Synthesis of N-Acyl-aminoacyl-pCpA for Preparation of Mischarged Fully Ribo tRNA

Electrophilic RNA for Peptidyl‐RNA Synthesis and Site‐Specific Cross‐Linking with tRNA‐Binding Enzymes

Electrophilic RNA for Peptidyl‐RNA Synthesis and Site‐Specific Cross‐Linking with tRNA‐Binding Enzymes

Contact Info

Product

Resources

About