Resolving Chemical Modifications to a Single Amino Acid within a Peptide Using a Biological NanoporeRestrepo-Pérez, L., Huang, G., Bohländer, P. R., Worp, N., Eelkema, R., Maglia, G., Joo, C. & Dekker, C., 24-Dec-2019, In : Acs Nano. 13, 12, p. 13668-13676 9 p.
Research output: Contribution to journal › Article › Academic › peer-review
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.
|Number of pages||9|
|Early online date||19-Sep-2019|
|Publication status||Published - 24-Dec-2019|
- nanopore, protein fingerprinting, amino acid labeling, protein analysis, biological nanopores, single-molecule protein sequencing, PROTEIN, GENOMICS, OMICS