TY - JOUR
T1 - Quantification of Protein Glycosylation Using Nanopores
AU - Versloot, Roderick Corstiaan Abraham
AU - Lucas, Florian Leonardus Rudolfus
AU - Yakovlieva, Liubov
AU - Tadema, Matthijs Jonathan
AU - Zhang, Yurui
AU - Wood, Thomas M
AU - Martin, Nathaniel I
AU - Marrink, Siewert J
AU - Walvoort, Marthe T C
AU - Maglia, Giovanni
PY - 2022/7/13
Y1 - 2022/7/13
N2 - Although nanopores can be used for single-molecule sequencing of nucleic acids using low-cost portable devices, the characterization of proteins and their modifications has yet to be established. Here, we show that hydrophilic or glycosylated peptides translocate too quickly across FraC nanopores to be recognized. However, high ionic strengths (i.e., 3 M LiCl) and low pH (i.e., pH 3) together with using a nanopore with a phenylalanine at its constriction allows the recognition of hydrophilic peptides, and to distinguish between mono- and diglycosylated peptides. Using these conditions, we devise a nanopore method to detect, characterize, and quantify post-translational modifications in generic proteins, which is one of the pressing challenges in proteomic analysis.
AB - Although nanopores can be used for single-molecule sequencing of nucleic acids using low-cost portable devices, the characterization of proteins and their modifications has yet to be established. Here, we show that hydrophilic or glycosylated peptides translocate too quickly across FraC nanopores to be recognized. However, high ionic strengths (i.e., 3 M LiCl) and low pH (i.e., pH 3) together with using a nanopore with a phenylalanine at its constriction allows the recognition of hydrophilic peptides, and to distinguish between mono- and diglycosylated peptides. Using these conditions, we devise a nanopore method to detect, characterize, and quantify post-translational modifications in generic proteins, which is one of the pressing challenges in proteomic analysis.
KW - protein glycosylation
KW - single molecule
KW - nanopore spectrometry
KW - rhamnosylation
KW - proteomics
U2 - 10.1021/acs.nanolett.2c01338
DO - 10.1021/acs.nanolett.2c01338
M3 - Article
C2 - 35766994
SN - 1530-6984
VL - 22
SP - 5357
EP - 5364
JO - Nano Letters
JF - Nano Letters
IS - 13
M1 - 2c10338
ER -