Stepwise Solid‐Phase Synthesis of Nucleopeptides

Grandas, Anna; Marchán, Vicente; Debéthune, Laurent; Pedroso, Enrique

doi:10.1002/0471142700.nc0422s31

Cited by 7 publications

(6 citation statements)

References 53 publications

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“…The syntheses were performed on a 10-mol scale according to established methods (22). Elongation was performed using 3Ј-phosphoramidite derivatives of DMT-protected nucleosides 5Ј-DMT-A(TAC)-OH, 5Ј-DMT-C(TAC)-OH, 5Ј-DMT-G(TAC)-OH, and 5Ј-DMT-T-OH under the agency of dicyanoimidazole.…”

Section: Oligonucleotide Synthesismentioning

confidence: 99%

Distinct Uptake Mechanisms but Similar Intracellular Processing of Two Different Toll-like Receptor Ligand-Peptide Conjugates in Dendritic Cells

Khan

Bijker

Weterings

et al. 2007

Journal of Biological Chemistry

163

139

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Covalent conjugation of Toll-like receptor ligands (TLR-L) to synthetic antigenic peptides strongly improves antigen presentation in vitro and T lymphocyte priming in vivo. These molecularly well defined TLR-L-peptide conjugates, constitute an attractive vaccination modality, sharing the peptide antigen and a defined adjuvant in one single molecule. We have analyzed the intracellular trafficking and processing of two TLR-L conjugates in dendritic cells (DCs). Long synthetic peptides containing an ovalbumin cytotoxic T-cell epitope were chemically conjugated to two different TLR-Ls the TLR2 ligand, Pam 3 CysSK 4 (Pam) or the TLR9 ligand CpG. Rapid and enhanced uptake of both types of TLR-L-conjugated peptide occurred in DCs. Moreover, TLR-L conjugation greatly enhanced antigen presentation, a process that was dependent on endosomal acidification, proteasomal cleavage, and TAP translocation. The uptake of the CpGϳ conjugate was independent of endosomally-expressed TLR9 as reported previously. Unexpectedly, we found that Pamϳconjugated peptides were likewise internalized independently of the expression of cell surface-expressed TLR2. Further characterization of the uptake mechanisms revealed that TLR2-L employed a different uptake route than TLR9-L. Inhibition of clathrin-or caveolindependent endocytosis greatly reduced uptake and antigen presentation of the Pam-conjugate. In contrast, internalization and antigen presentation of CpGϳconjugates was independent of clathrin-coated pits but partly dependent on caveolae formation. Importantly, in contrast to the TLR-independent uptake of the conjugates, TLR expression and downstream TLR signaling was required for dendritic cell maturation and for priming of naïve CD8؉ T-cells. Together, our data show that targeting to two distinct TLRs requires distinct uptake mechanism but follows similar trafficking and intracellular processing pathways leading to optimal antigen presentation and T-cell priming.

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Section: Oligonucleotide Synthesismentioning

confidence: 99%

Distinct Uptake Mechanisms but Similar Intracellular Processing of Two Different Toll-like Receptor Ligand-Peptide Conjugates in Dendritic Cells

Khan

Bijker

Weterings

et al. 2007

Journal of Biological Chemistry

163

139

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“…Therefore, finding the correct protecting group compatible with both oligonucleotide and peptide moieties is the main challenge of solid-phase synthesis. Besides, only amino acids with no reactive side chains like alanine and leucine or those with easily removable side chain protection groups can be conjugated with oligonucleotides via the solid phase method [ 54 , 65 ]. At present, no POC introduced as an ss-aRNase has been prepared by solid phase methods.…”

Section: Synthetic Approaches Applied For the Generation Of Site-smentioning

confidence: 99%

Site-Selective Artificial Ribonucleases: Renaissance of Oligonucleotide Conjugates for Irreversible Cleavage of RNA Sequences

et al. 2021

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RNA-targeting therapeutics require highly efficient sequence-specific devices capable of RNA irreversible degradation in vivo. The most developed methods of sequence-specific RNA cleavage, such as siRNA or antisense oligonucleotides (ASO), are currently based on recruitment of either intracellular multi-protein complexes or enzymes, leaving alternative approaches (e.g., ribozymes and DNAzymes) far behind. Recently, site-selective artificial ribonucleases combining the oligonucleotide recognition motifs (or their structural analogues) and catalytically active groups in a single molecular scaffold have been proven to be a great competitor to siRNA and ASO. Using the most efficient catalytic groups, utilising both metal ion-dependent (Cu(II)-2,9-dimethylphenanthroline) and metal ion-free (Tris(2-aminobenzimidazole)) on the one hand and PNA as an RNA recognising oligonucleotide on the other, allowed site-selective artificial RNases to be created with half-lives of 0.5–1 h. Artificial RNases based on the catalytic peptide [(ArgLeu)2Gly]2 were able to take progress a step further by demonstrating an ability to cleave miRNA-21 in tumour cells and provide a significant reduction of tumour growth in mice.

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“…Monofunctionalized Linkers. Grandas' research group has long been involved in the syntheses of various nucleopeptides . In some of the natural nucleopeptides, the side chain hydroxyl group of the peptides containing serine, threonine, or tyrosine is linked to the 5‘-terminus of the oligonucleotide by a phosphonate diester linkage, as in Chart (scP→5‘).…”

Section: 11 Solid-phase-immobilized Linkers Used In Sequential Synthe...mentioning

confidence: 99%

Peptide Conjugates of Oligonucleotides: Synthesis and Applications

2006

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Stepwise Solid‐Phase Synthesis of Nucleopeptides

Cited by 7 publications

References 53 publications

Distinct Uptake Mechanisms but Similar Intracellular Processing of Two Different Toll-like Receptor Ligand-Peptide Conjugates in Dendritic Cells

Distinct Uptake Mechanisms but Similar Intracellular Processing of Two Different Toll-like Receptor Ligand-Peptide Conjugates in Dendritic Cells

Site-Selective Artificial Ribonucleases: Renaissance of Oligonucleotide Conjugates for Irreversible Cleavage of RNA Sequences

Peptide Conjugates of Oligonucleotides: Synthesis and Applications

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