Identification of Disulfide Bonds in Protein Proteolytic Degradation Products Using de novo-Protein Unique Sequence Tags Approach
peer-reviewed · Journal of Proteome Research · 2010
| Date | 2010-08-06 |
| Type | peer-reviewed |
| Venue | Journal of Proteome Research |
| Publisher | American Chemical Society (ACS) |
| Contribution | algorithm |
| DOI | 10.1021/pr1002559 |
| Citations (OpenAlex) | 7 |
| Venue 2-year citedness | 3.83 |
Abstract
Disulfide bonds are a form of post-translational modification that often determines protein structure(s) and function(s). In this work, we report a mass spectrometry method for identification of disulfides in degradation products of proteins, specifically endogenous peptides in the human blood plasma peptidome. LC-Fourier transform tandem mass spectrometry (FT MS/MS) was used for acquiring mass spectra that were de novo sequenced and then searched against the IPI human protein database. Through the use of unique sequence tags (UStags), we unambiguously correlated the spectra to specific database proteins. Examination of the UStags’ prefix and/or suffix sequences that contain cysteine(s) in conjunction with sequences of the UStags-specified database proteins is shown to enable the unambigious determination of disulfide bonds. Using this method, we identified the intermolecular and intramolecular disulfides in human blood plasma peptidome peptides that have molecular weights of up to approximately 10 kDa.
Methods and tools
- Unique sequence tags for PTM discovery: Reads short, uniquely assignable sequence tags de novo and uses them to find post-translational modifications, trading the full-length sequence for the reliability that made early de novo output hard to trust in routine proteomics.