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Label-Free Detection of Post-translational Modifications with a Nanopore
journal contribution
posted on 2019-10-17, 20:14 authored by Laura Restrepo-Pérez, Chun Heung Wong, Giovanni Maglia, Cees Dekker, Chirlmin JooPost-translational
modifications (PTMs) of proteins play key roles
in cellular processes. Hence, PTM identification is crucial for elucidating
the mechanism of complex cellular processes and disease. Here we present
a method for PTM detection at the single-molecule level using FraC
biological nanopores. We focus on two major PTMs, phosphorylation
and glycosylation, that mutually compete for protein modification
sites, an important regulatory process that has been implicated in
the pathogenic pathways of many diseases. We show that phosphorylated
and glycosylated peptides can be clearly differentiated from nonmodified
peptides by differences in the relative current blockade and dwell
time in nanopore translocations. Furthermore, we show that these PTM
modifications can be mutually differentiated, demonstrating the identification
of phosphorylation and glycosylation in a label-free manner. The results
represent an important step for the single-molecule, label-free identification
of proteoforms, which have tremendous potential for disease diagnosis
and cell biology.
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protein modification sitesPTM identificationcell biologyglycosylationnanopore translocationssingle-molecule levelPTM modificationsPost-translational Modificationsnonmodified peptidesPTM detectionlabel-free identificationLabel-Free DetectionphosphorylationNanopore Post-translational modificationslabel-free mannerglycosylated peptidesdisease diagnosis
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