TY - JOUR
T1 - Resolving Chemical Modifications to a Single Amino Acid within a Peptide Using a Biological Nanopore
AU - Restrepo-Pérez, Laura
AU - Huang, Gang
AU - Bohländer, Peggy R.
AU - Worp, Nathalie
AU - Eelkema, Rienk
AU - Maglia, Giovanni
AU - Joo, Chirlmin
AU - Dekker, Cees
N1 - Publisher Copyright:
© 2019 American Chemical Society.
PY - 2019/12/24
Y1 - 2019/12/24
N2 - While DNA sequencing is now amply available, fast, and inexpensive, protein sequencing remains a tremendous challenge. Nanopores may allow for developing a protein sequencer with single-molecule capabilities. As identification of 20 different amino acids currently presents an unsurmountable challenge, fingerprinting schemes are pursued, in which only a subset of amino acids is labeled and detected. This requires modification of amino acids with chemical structures that generate a distinct nanopore ionic current signal. Here, we use a model peptide and the fragaceatoxin C nanopore to characterize six potential tags for a fingerprinting approach using nanopores. We find that labeled and unlabeled proteins can be clearly distinguished and that sensitive detection is obtained for labels with a spectrum of different physicochemical properties such as mass (427-1275 Da), geometry, charge, and hydrophobicity. Additionally, information about the position of the label along the peptide chain can be obtained from individual current-blockade event features. The results represent an important advance toward the development of a single-molecule protein-fingerprinting device with nanopores.
AB - While DNA sequencing is now amply available, fast, and inexpensive, protein sequencing remains a tremendous challenge. Nanopores may allow for developing a protein sequencer with single-molecule capabilities. As identification of 20 different amino acids currently presents an unsurmountable challenge, fingerprinting schemes are pursued, in which only a subset of amino acids is labeled and detected. This requires modification of amino acids with chemical structures that generate a distinct nanopore ionic current signal. Here, we use a model peptide and the fragaceatoxin C nanopore to characterize six potential tags for a fingerprinting approach using nanopores. We find that labeled and unlabeled proteins can be clearly distinguished and that sensitive detection is obtained for labels with a spectrum of different physicochemical properties such as mass (427-1275 Da), geometry, charge, and hydrophobicity. Additionally, information about the position of the label along the peptide chain can be obtained from individual current-blockade event features. The results represent an important advance toward the development of a single-molecule protein-fingerprinting device with nanopores.
KW - amino acid labeling
KW - biological nanopores
KW - nanopore
KW - protein analysis
KW - protein fingerprinting
KW - single-molecule protein sequencing
UR - http://www.scopus.com/inward/record.url?scp=85073007805&partnerID=8YFLogxK
U2 - 10.1021/acsnano.9b05156
DO - 10.1021/acsnano.9b05156
M3 - Article
C2 - 31536327
AN - SCOPUS:85073007805
SN - 1936-0851
VL - 13
SP - 13668
EP - 13676
JO - ACS Nano
JF - ACS Nano
IS - 12
ER -