Single-molecule peptide sequencing through reverse translation of peptides into DNA - PubMed
3 hours ago
- #single-molecule
- #protein sequencing
- #DNA barcoding
- Single-molecule peptide sequencing is achieved through reverse translation of peptides into DNA.
- The method involves a modified Edman degradation process that releases N-terminal amino acids tagged with DNA barcodes.
- Antibody-mediated proximity extension assays identify barcoded amino acids, generating PCR-amplifiable DNA reporters.
- High-throughput sequencing reads the DNA library, converting peptide sequences into digital DNA outputs.
- The approach provides full sequence coverage in millions of reads and differentiates native and post-translationally modified peptides.
- This framework redefines protein sequencing as a DNA sequencing problem, enabling high-throughput, de novo single-molecule protein sequencing.