Biomolecular characterization of allergenic proteins in snow crab (Chionoecetes opilio) and de novo sequencing of the second allergen arginine kinase using tandem mass spectrometry.
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.
Department of Chemistry, Memorial University of Newfoundland, St. John's, Newfoundland and Labrador, A1B 3X7, Canada. anasar@mun.ca
Anas M
AM
Abdel Rahman
Sandip D
SD
Kamath
Andreas L
AL
Lopata
John J
JJ
Robinson
Robert J
RJ
Helleur
D000485:Allergens / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D001122:Arginine Kinase / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D003386:Brachyura / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009124:Muscle Proteins / Q000032:analysis / Q000737:chemistry / Q000276:immunology; D011506:Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry
D000485:Allergens; D009124:Muscle Proteins; D011506:Proteins; D001122:Arginine Kinase