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Detection of phosphorylation post-translational modifications along single peptides with nanopores

Nat Biotechnol. 2023-06; 
Ian C Nova, Justas Ritmejeris, Henry Brinkerhoff, Theo J R Koenig, Jens H Gundlach, Cees Dekker
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Peptide Synthesis … DNA oligo (the ‘threading DNA’) to the other end of the peptide 16 (Fig. 1a). This DNA … -28b(+) vector plasmid at Ndel/NotI sites by Genscript. Ion current data were acquired at 50 kHz … Get A Quote

摘要

Current methods to detect post-translational modifications of proteins, such as phosphate groups, cannot measure single molecules or differentiate between closely spaced phosphorylation sites. We detect post-translational modifications at the single-molecule level on immunopeptide sequences with cancer-associated phosphate variants by controllably drawing the peptide through the sensing region of a nanopore. We discriminate peptide sequences with one or two closely spaced phosphates with 95% accuracy for individual reads of single molecules.

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