Biomolecular characterization of allergenic proteins in snow crab (Chionoecetes opilio) and de novo sequencing of the second allergen arginine kinase using tandem mass spectrometry
peer-reviewed · Journal of Proteomics · 2011
| Date | 2011-02-01 |
| Type | peer-reviewed |
| Venue | Journal of Proteomics |
| Publisher | Elsevier BV |
| Contribution | downstream-application |
| DOI | 10.1016/j.jprot.2010.10.010 |
| Citations (OpenAlex) | 88 |
| Venue 2-year citedness | 3.02 |
Abstract
Snow crab (Chionoecetes opilio) proteins have been recognized as an important source of both food and occupational allergens. While snow crab causes a significant occupational allergy, only one novel allergen has recently been fully characterized. The muscle proteins from snow crab legs were profiled by SDS-PAGE. Several of these proteins were characterized using tandem mass spectrometry. Five proteins were identified; sarcoplasmic Ca-binding (20kDa), arginine kinase (40), troponin (23kDa) and α-actine (42kDa) and smooth endoplasmic reticulum Ca(2+)ATPase (113kDa). Immunoblotting using serum of sixteen allergic patients resulted in strong reactivity with the 40-kDa protein in seven patients (43%). This protein was purified by chromatography and subsequently de novo sequenced using matrix assisted laser desorption ionization and electrospray tandem mass spectrometry. We identified a second important allergen, arginine kinase, in snow crab, designated Chi o 3. Based on identity and homology analysis, using bioinformatics tools, a signature peptide was identified as a chemical surrogate for arginine kinase. The suitability of this signature peptide was tested for analytically representing the arginine kinase, by performing a multi-reaction monitoring tandem mass spectrometry approach on actual air filter samples collected from a simulated crab processing plant.
Methods and tools
- Snow crab allergen de novo sequencing: Profiled snow crab muscle allergens and de novo sequenced the newly recognised 40 kDa allergen, arginine kinase, by MALDI and ESI tandem mass spectrometry; the group also de novo sequenced its tropomyosin.
Cites (2)
- Characterization and de novo sequencing of snow crab tropomyosin enzymatic peptides by both electrospary ionization and matrix‐assisted laser desorption ionization QqToF tandem mass spectrometry (2010) crossref
- De novo sequence analysis of N‐terminal sulfonated peptides after in‐gel guanidination (2005) crossref