Fig. 1: Covalent RaPID setup. | Communications Chemistry

Fig. 1: Covalent RaPID setup.

From: Discovering covalent cyclic peptide inhibitors of peptidyl arginine deiminase 4 (PADI4) using mRNA-display with a genetically encoded electrophilic warhead

Fig. 1

A The structure of arginine and related arginine-mimetic PADI4 inhibitor 1 and synthesised unnatural amino acids 2 and 3. B Microhelix assay using a truncated tRNA mimic to monitor loading of FAO-CBT (3) using eFx. The upper band indicates the presence of aminoacylated microhelix RNA and the lower band is non-aminoacylated microhelix RNA. After 2 h incubation at 4 °C, 59% aminoacylation is seen. The full gel is provided in Fig. S23. C The covalent RaPID cycle setup. Transcription, puromycin ligation, translation, reverse transcription, and affinity panning against immobilised PADI4 are performed as in a typical RaPID selection. However, denaturing washes are added as an additional step to remove non-covalent peptide binders to PADI4. The translation incorporates FAO (2) and chloroacetylated-D-tyrosine to promote covalent binding and cyclisation, respectively.

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