Development of a Novel Peptide Aptamer that Interacts with the eIF4E Capped-mRNA Binding Site using Peptide Epitope Linker Evolution (PELE).

Frosi, Y.; Ng, S.; Lin, Y.-C.; Jiang, S.; Ramlan, S. R.; Lama, D.; Verma, C. S.; Asial, I.; Brown, C. J.

doi:10.1101/2022.02.17.480295

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bioRxiv · 10.1101/2022.02.17.480295

Development of a Novel Peptide Aptamer that Interacts with the eIF4E Capped-mRNA Binding Site using Peptide Epitope Linker Evolution (PELE).

Frosi, Y.·Ng, S.·Lin, Y.-C.·Jiang, S.·Ramlan, S. R.·Lama, D.·Verma, C. S.·Asial, I.·Brown, C. J.

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Abstract

Identifying new binding sites and poses that modify biological function are an important step towards drug discovery. We have identified a novel disulphide constrained peptide that interacts with the cap-binding site of eIF4E, an attractive therapeutic target that is commonly overexpressed in many cancers and plays a significant role in initiating a cancer specific protein synthesis program though binding the 5cap (7methyl-guanoisine) moiety found on mammalian mRNAs. The use of disulphide constrained peptides to explore intracellular biological targets is limited by their lack of cell permeability and the instability of the disulphide bond in the reducing environment of the cell, loss of which results in abrogation of binding. To overcome these challenges, the cap-binding site interaction motif was placed in a hypervariable loop on an VH domain, and then selections performed to select a molecule that could recapitulate the interaction of the peptide with the target of interest in a process termed Peptide Epitope Linker Evolution (PELE). A novel VH domain was identified that interacted with the eIF4E cap binding site with a nanomolar affinity and that could be intracellularly expressed in mammalian cells. Additionally, it was demonstrated to specifically modulate eIF4E function by decreasing cap-dependent translation and cyclin D1 expression, common effects of eIF4F complex disruption.

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Frosi, Y., Ng, S., Lin, Y.-C., Jiang, S., Ramlan, S. R., Lama, D., Verma, C. S., Asial, I., Brown, C. J.. 2022-02-18. Development of a Novel Peptide Aptamer that Interacts with the eIF4E Capped-mRNA Binding Site using Peptide Epitope Linker Evolution (PELE).. https://doi.org/10.1101/2022.02.17.480295

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