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

peer-reviewed · Journal of Mass Spectrometry · 2010

peer-reviewed · Journal of Mass Spectrometry · 2010. Anas M. Abdel Rahman et al. The protein tropomyosin (TM) is a known major allergen present in shellfish causing frequent food allergies…
Date 2010-04-01
Type peer-reviewed
Venue Journal of Mass Spectrometry
Publisher Wiley
Contribution downstream-application
DOI 10.1002/jms.1721
Citations (OpenAlex) 38
Venue 2-year citedness 1.85

Abstract

The protein tropomyosin (TM) is a known major allergen present in shellfish causing frequent food allergies. TM is also an occupational allergen generated in the working environment of snow crab (Chionoecetes opilio) processing plants. The TM protein was purified from both claw and leg meats of snow crab and analyzed by electrospray ionization and matrix-assisted laser desorption/ionization (MALDI) using hybrid quadruple time-of-flight tandem mass spectrometry (QqToF-MS). The native polypeptide molecular weight of TM was determined to be 32,733 Da. The protein was further characterized using the ‘bottom-up’ MS approach. A peptide mass fingerprinting was obtained by two different enzymatic digestions and de novo sequencing of the most abundant peptides performed. Any post-translational modifications were identified by searching their calculated and predicted molecular weights in precursor ion spectra. The immunological reactivity of snow crab extract was evaluated using specific antibodies and allergenic reactivity assessed with serum of allergic patients. Subsequently, a signature peptide for TM was identified and evaluated in terms of identity and homology using the basic local alignment search tool (BLAST). The identification of a signature peptide for the allergen TM using MALDI-QqToF-MS will be critical for the sensitive and specific quantification of this highly allergenic protein in the work place.

Authors

  1. Anas M. Abdel Rahman · Memorial University of Newfoundland
  2. Andreas L. Lopata · Australian Institute of Tropical Health and Medicine, James Cook University, RMIT University
  3. Robyn E. O’Hehir · Monash University, The Alfred Hospital
  4. John J. Robinson · Memorial University of Newfoundland
  5. Joseph Banoub · Fisheries and Oceans Canada, Memorial University of Newfoundland
  6. Robert J. Helleur · Memorial University of Newfoundland

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 (3)

Cited by (1)

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