期刊
ACS NANO
卷 13, 期 12, 页码 13668-13676出版社
AMER CHEMICAL SOC
DOI: 10.1021/acsnano.9b05156
关键词
nanopore; protein fingerprinting; amino acid labeling; protein analysis; biological nanopores; single-molecule protein sequencing
类别
资金
- European Research Council Advanced Grant SynDiv [669598]
- Netherlands Organisation for Scientific Research (NWO/OCW) through the NanoFront grant
- Netherlands Organisation for Scientific Research (NWO/OCW) through the BaSyC grant
- Foundation for Fundamental Research on Matter (Vrije Programma SMPS)
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.
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