TY - JOUR
T1 - Label-Free Detection of Post-translational Modifications with a Nanopore
AU - Restrepo-Pérez, Laura
AU - Wong, Chun Heung
AU - Maglia, Giovanni
AU - Dekker, Cees
AU - Joo, Chirlmin
PY - 2019
Y1 - 2019
N2 - Post-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.
AB - Post-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.
KW - glycosylation
KW - label-free detection
KW - Nanopores
KW - phosphorylation
KW - post-translational modifications
UR - http://www.scopus.com/inward/record.url?scp=85073880659&partnerID=8YFLogxK
U2 - 10.1021/acs.nanolett.9b03134
DO - 10.1021/acs.nanolett.9b03134
M3 - Article
AN - SCOPUS:85073880659
VL - 19
SP - 7957
EP - 7964
JO - Nano Letters: a journal dedicated to nanoscience and nanotechnology
JF - Nano Letters: a journal dedicated to nanoscience and nanotechnology
SN - 1530-6984
IS - 11
ER -