Label-Free Detection of Post-translational Modifications with a Nanopore

Label-Free Detection of Post-translational Modifications with a Nanopore

Restrepo Perez, L. (TU Delft BN/Chirlmin Joo Lab; Kavli institute of nanoscience Delft)
Wong, C.H. (TU Delft BUS/Quantum Delft; Kavli institute of nanoscience Delft)
Maglia, Giovanni (Rijksuniversiteit Groningen)
Dekker, C. (TU Delft BN/Cees Dekker Lab; Kavli institute of nanoscience Delft)
Joo, C. (TU Delft BN/Chirlmin Joo Lab; Kavli institute of nanoscience Delft)

2019

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.

glycosylation
label-free detection
Nanopores
phosphorylation
post-translational modifications

http://resolver.tudelft.nl/uuid:29f98b57-998f-4f6f-ba8e-660f44b42e45

https://doi.org/10.1021/acs.nanolett.9b03134

1530-6984

Nano Letters: a journal dedicated to nanoscience and nanotechnology, 19 (11), 7957-7964

Institutional Repository

journal article

© 2019 L. Restrepo Perez, C.H. Wong, Giovanni Maglia, C. Dekker, C. Joo