Open AccessQuantification of Protein Glycosylation Using Nanopores
Author(s) -
Roderick Corstiaan Abraham Versloot,
Florian Leonardus Rudolfus Lucas,
Liubov Yakovlieva,
Matthijs Jonathan Tadema,
Yurui Zhang,
Thomas M. Wood,
Nathaniel I. Martin,
Siewert J. Marrink,
Marthe T. C. Walvoort,
Giovanni Maglia
Publication year - 2022
Publication title -
nano letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 4.853
H-Index - 488
eISSN - 1530-6992
pISSN - 1530-6984
DOI - 10.1021/acs.nanolett.2c01338
Subject(s) - nanopore , chemistry , glycosylation , nucleic acid , nanotechnology , nanopore sequencing , ionic bonding , biophysics , amino acid , peptide , biochemistry , materials science , biology , ion , dna , organic chemistry , dna sequencing
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.