Department of Chemistry, Memorial University of Newfoundland, St. John's, Newfoundland, A1B 3X7, Canada.
Rapid Commun Mass Spectrom. 2010 Aug 30;24(16):2462-70. doi: 10.1002/rcm.4664.
Crustaceans are the third most prevalent cause of food-induced anaphylaxis after peanuts and tree nuts. The severity of the allergenic proteins depends mainly on the amino acid sequence that induces production of IgE antibodies. In black tiger prawn (Penaeus monodon), the crude protein extract was profiled and its allergenic potency was examined against patient's sera. Proteins having strong immunoreactivity with patient's IgE were characterized using peptide mass fingerprinting (PMF). Tropomyosin (TM) (33 kDa), myosin light chain (20 kDa), and arginine kinase (40 kDa) were identified as allergenic proteins. Tropomyosin, the most abundant and potent allergen, was purified using ion-exchange chromatography for de novo sequencing experiments. Using bottom up tandem mass spectrometry, the full amino acid sequence was achieved by a combination of matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) tandem mass spectrometry (QqToF). Myosin light chain and arginine kinase were also characterized, and their related peptides were de novo sequenced using the same approach. The immunological reactivity of the crude prawn extracts and purified TM samples were analyzed using a large number of patients' sera. A signature peptide was assigned for the TM protein for future quantification work of black tiger prawn TM levels in different matrices (i.e. water, air, food) in the seafood industry.
甲壳类动物是继花生和坚果之后引起食物过敏的第三大过敏原。过敏原蛋白的严重程度主要取决于诱导 IgE 抗体产生的氨基酸序列。在黑虎虾（Penaeus monodon）中，对粗蛋白提取物进行了分析，并针对患者的血清检查了其过敏原效价。使用肽质量指纹图谱（PMF）对与患者 IgE 具有强免疫反应性的蛋白质进行了表征。肌球蛋白轻链（20 kDa）和精氨酸激酶（40 kDa）被鉴定为过敏原蛋白。肌球蛋白轻链和精氨酸激酶也被表征，并使用相同的方法对其相关肽进行从头测序。使用基质辅助激光解吸/电离（MALDI）和电喷雾电离（ESI）串联质谱（QqToF）组合，通过自上而下的串联质谱法获得了全长氨基酸序列。使用大量患者的血清分析了粗虾提取物和纯化 TM 样品的免疫反应性。为了在未来对不同基质（即水、空气、食物）中黑虎虾 TM 水平进行定量工作，为 TM 蛋白分配了一个特征肽。
