Comprehensive Proteomics Approach in Characterizing and Quantifying Allergenic Proteins from Northern Shrimp: Toward Better Occupational Asthma Prevention

Abstract

Occupational asthma is a major chronic health dilemma among workers involved in the seafood industry. Several proteins notoriously known to cause asthma have been reported in different seafood. This work involves the application of an allergenomics strategy to study the most potent allergens of northern shrimp. The proteins were extracted from shrimp tissue and profiled by gel electrophoresis. Allergenic proteins were identified based on their reactivity to patient sera and were structurally identified using tandem mass spectrometry. Northern shrimp tropomyosin, arginine kinase, and sarcoplasmic calcium-binding protein were found to be the most significant allergens. Multiple proteolytic enzymes enabled 100% coverage of the sequence of shrimp tropomyosin by tandem mass specrometry. Only partial sequence coverage was obtained, however, for the shrimp allergen arginine kinase. Signature peptides, for both tropomyosin and arginine kinase, were assigned and synthesized for use in developing the multiple reaction monitoring tandem mass spectrometric method. Subsequently, air samples were collected from a shrimp processing plant and two aerosolized proteins quantified using tandem mass specrometry. Allergens were detected in all areas of the plant, reaching levels as high as 375 and 480 ng/m(3) for tropomyosine and arginine kinase, respectively. Tropomyosine is much more abundant than arginine kinase in shrimp tissues, so the high levels of arginine kinase suggest it is more easily aerosolized. The present study shows that mass spectrometric analysis is a sensitive and accurate tool in identifying and quantifying aerosolized allergens.

Effect of heat processing on antibody reactivity to allergen variants and fragments of black tiger prawn: A comprehensive allergenomic approach

“…Interestingly, after heat treatment, AK IgE reactivity was reduced to just one AK variant at 36.5 kDa. We recently demonstrated that the heat‐stable novel AK from northern shrimp can be aerosolized and cause inhalational sensitization in the workplace .…”

Shotgun proteomics, in-silico evaluation and immunoblotting assays for allergenicity assessment of lesser mealworm, black soldier fly and their protein hydrolysates

“…Grinded insects were defatted with diethyl ether following the method proposed by Caligiani et al 31 using a Soxhlet extractor (SER 148/3 VELP SCIENTIFICA, Usmate Velate, Italy). Defatted flours were subjected to the protein extraction protocol proposed by Abdel Rahman et al 32 with some modifications. Briefly, 0.5 g of sample were homogenized in an ice-bath with 10 mL of a Tris buffer (25 mM Tris-HCl pH 8.0, 1 M KCl, 50 mM DTT and 0.5 mM EDTA) using Ultra Turrax homogenizer (11000 rpm) for 1 minute.…”

Shellfish allergens: tropomyosin and beyond

“…Troponin C (TpC) was first described as an allergen in Crangon crangon (Cra c 6, North Sea shrimp) , recognized by 9/31 (29%) of the patients, and later in Pandalus borealis (northern shrimp), recognized by 33% of the patients . In a recent study by Pascal et al , sensitization to TpC was found in 17.2% of 58 shrimp‐allergic patients.…”

“…During the last two decades, significant progress in biochemistry and molecular biology enabled the characterization, cloning, and recombinant production of various shellfish allergen components and epitope‐emulating peptides that might become available for quantification of specific IgE (sIgE) antibodies, namely molecular diagnosis. This review intends to summarize our current knowledge about shellfish allergens and their cross‐reactivity as this might be the key to optimize diagnosis .…”