Identification of Major and Minor Allergens of Mud Crab (Scylla Serrata)
Identification of Mud Crab Allergens
Vol. 10 No. 2 : 2015 (90-97)
Abstract
Crab meat is a valuable source of proteins and functional lipids and is widely consumed worldwide. However, the prevalence of crab allergy has increased over the past few years. In order to better understand crab allergy, it is necessary to identify crab allergens. The aim of the present study was to compare the IgE-binding proteins of raw and cooked extracts of mud crab (Scylla serrata). Raw and cooked extracts of the mud crab were prepared. Protein profiles and IgE reactivity patterns were identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) followed by immunoblotting using sera from 21 skin prick test (SPT) positive patients. In SDS-PAGE, 20 protein bands (12 to 250 kDa) were observed in the raw extract while the cooked extract demonstrated fewer bands. Protein bands between 40 to 250 kDa were sensitive to heat denaturation and no longer observed in the cooked extract. In immunoblotting experiments, raw and cooked extracts demonstrated 11 and 4 IgE-binding proteins, respectively, with molecular weights of between 23 and 250 kDa. A heat-resistant 36 kDa protein, corresponding to crab tropomyosin was identified as the major allergen of both extracts. In addition, a 41 kDa heat-sensitive protein believed to be arginine kinase was shown to be a major allergen of the raw extract. Other minor allergens were also observed at various molecular weights.
Keywords :
arginine kinase,
immunoblotting,
Scylla serrata,
tropomyosin,
Correspondance Address
Nurul Izzah Abdul Rahman, Allergy and Immunology Research Centre, Institute for Medical Research, Jalan Pahang, 50588 Kuala Lumpur, Malaysia. Tel: 03-26162789 Fax: 03-26912019 Email: nurulizzah012@gmail.com
References
Abdel Rahman, A.M., Kamath, S.D., Lopata, A.L., Robinson, J.J., Helleur, R.J. 2011. Biomolecular characterization of allergenic proteins in snow crab (Chionoecetes opilio) and de novo sequencing of the second allergen arginine kinase using tandem mass spectrometry. J Proteomics 74(2): 231-41.
Abramovitch, J.B., Kamath, S., Varese, N., Zubrinich, C., Lopata, A.L., O’Hehir, R.E., Rolland, J.M. 2013. IgE reactivity of blue swimmer crab (portunus pelagicus) tropomyosin, por p 1, and other allergens; cross-reactivity with black tiger prawn and effects of heating. PLoS One 8(6): e67487.
Ayuso, R., Reese, G., Leong-Kee, S., Plante, M., Lehrer, S.B. 2002. Molecular basis of arthropod cross-reactivity: IgE-binding cross-reactive epitopes of shrimp, house dust mite and cockroach tropomyosins. Int Arch Allergy Immunol 129(1): 38-48.
Besler, M., Steinhart, H., Paschke, A. 2001. Stability of food allergens and allergenicity of processed food. J Chromatogr B Biomed Sci Appl 756(1-2): 207-28.
Butt, C., Macdougall, C. 2008. Diagnosing and managing food allergy in children. Paediatrics and Child Health 18(7): 317-22.
Chu, K.H., Wong, S.H., Leung, P.S. 2000. Tropomyosin is the major mollusk allergen: reverse transcriptase polymerase chain reaction, expression and ige reactivity. Mar Biotechnol (NY) 2(5): 499-509.
Fontan, M., Anibarro, B., Postigo, I., Martinez, J. 2005. Allergy to freshwater shrimp (Gammarus). J Investig Allergol Clin Immunol 15(2): 150-2.
Food and Agriculture Organization of the United Nations (FAO). 2010. FAO yearbook. fishery and aquaculture statistics 2010. Rome. http://www.fao.org/docrep/015/ba0058t/ba0058t.pdf [19 February 2015]
Hendrickx, M., Ludikhuyze, L., Van den broeck, I., Weemaes, C. 1998. Effect of high pressure on enzymes related to food quality. Trends in Food Science & Technology 9(5): 197-203.
Huang, Y.Y., Liu, G.M., Cai, Q.F., Weng, W.Y., Maleki, S.J., Su, W.J., Cao, M.J. 2010. Stability of major allergen tropomyosin and other food proteins of mud crab (Scylla serrata) by in vitro gastrointestinal digestion. Food Chem Toxicol 48(5): 1196-1201.
Hsu, M.L,, Li. L.F. 2012. Prevalence of food avoidance and food allergy in Chinese patients with chronic urticaria. Br J Dermatol 166(4): 747-52.
Keenan, C.P. 1999. Agriculture of the mud crab, genus Scylla - past, present, future. In A mud crab Agriculture and Biology, ACIAR Proceedings, No. 78. Edited by Keenan, C.P. and Blckshaw, A. Australia: Watson Ferguson and Company, Canberra; 9-13.
Lehrer, S.B., Ayuso, R., Reese, G. 2003. Seafood allergy and allergens: a review. Mar Biotechnol (NY) 5(4): 339-48.
Liang, Y., Cao, M., Su, W., Zhang, L., Huang, Y., Liu, G. 2008. Identification and characterization of the major allergens of Chinese mitten crab (Eriocheir sinensis). J Food Chem 111: 998-1003.
Liu, G.M., Cao, M.J., Huang, Y.Y., Cai, Q.F., Weng, W.Y., Su, W.J. 2010a. Comparative study of in vitro digestibility of major allergen tropomyosin and other food proteins of Chinese mitten crab (Eriocheir sinensis). J Sci Food Agric 90(10): 1614-20.
Liu, G.M., Cheng, H., Nesbit, J.B., Su, W.J., Cao, M.J., Maleki, S.J. 2010b. Effects of boiling on the IgE-binding properties of tropomyosin of shrimp (Litopenaeus vannamei). J Food Sci 75(1): T1-5.
Liu, G.M., Li, B., Yu, H.L., Cao, M.J., Cai, Q.F., Lin, J.W., Su, W.J. 2012. Induction of mud crab (Scylla paramamosain) tropomyosin and arginine kinase specific hypersensitivity in BALB/c mice. J Sci Food Agric 92(2): 232-8.
Lopata, A.L., Lehrer, S.B. 2009. New insights into seafood allergy. Curr Opin Allergy Clin Immunol 9(3): 270-7.
Ma, X.L., Sun, Q.M., Jia, J., Cai, L. 2012. Analysis of serum specific IgE in aeroallergen and food allergen in patients with allergic skin diseases. Beijing Da Xue Xue Bao 44(5): 765-9.
Nagpal, S., Rajappa, L., Metcalfe, D.D., Rao, P.V. 1989. Isolation and characterization of heat-stable allergens from shrimp (Penaeus indicus). J Allergy Clin Immunol 83(1): 26-36.
Peng, J., Gygi, S.P. 2001. Proteomics: the move to mixtures. J Mass Spectrom 36(10): 1083-91.
Reese, G., Ayuso, R., Lehrer, S.B. 1999. Tropomyosin: an invertebrate pan allergen. Int Arch Allergy Immunol 119(4): 247-58.
Rosmilah, M., Shahnaz, M., Zailatul, H.M., Noormalin, A., Normilah, I. 2012. Identification of tropomyosin and arginine kinase as major allergens of Portunus pelagicus (blue swimming crab). Trop Biomed 29(3): 467-78.
Sathe, S.K., Teuber, S.S., Roux, K.H. 2005. Effects of food processing on the stability of food allergens. Biotechnol Adv 23(6): 423-9.
Shahnaz, M., Gendeh, B.S., Nasuruddin, A. 2001. Skin test reactivity to inhalant food allergens in patients with allergic rhintis. Int Med Res J 5(2): 69–73.
Shen, Y., Cao, M.J., Cai, Q.F., Su, W.J., Yu, H.L., Ruan, W.W., Liu, G.M. 2011. Purification, cloning, expression and immunological analysis of Scylla serrata arginine kinase, the crab allergen. J Sci Food Agric 91(7): 1326-35.
Shriver, S.K., Yang, W. 2011. Thermal and nonthermal methods for food allergen control. Food Eng Rev 3: 26-43.
Suma, Y., Ishizaki, S., Nagashima, Y., Lu, Y., Ushio, H., Shiomi, K. 2007. Comparative analysis of barnacle tropomyosin: divergence from decapod tropomyosins and role as a potential allergen. Comp Biochem Physiol B Biochem Mol Biol 147(2): 230-6.
Yadzir, Z.H., Misnan, R., Abdullah, N., Bakhtiar, F., Arip, M., Murad, S. 2010. Characterization of IgE-binding proteins of raw and cooked extracts of Loligo edulis (white squid). Southeast Asian J Trop Med Public Health 41(3): 653-9.
Yu, C.J., Lin, Y.F., Chiang, B.L., Chow, L.P. 2003. Proteomics and immunological analysis of a novel shrimp allergen, Pen m 2. J Immunol 170(1): 445-53.
Yu, H.L., Ruan, W.W., Cao, M.J., Cai, Q.F., Shen, H.W., Liu, G.M. 2013. Identification of physicochemical properties of Scylla paramamosain allergen, arginin kinase. J Sci Food Agric 93(2): 245-53.
