UPSI Digital Repository (UDRep)
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Abstract : Universiti Pendidikan Sultan Idris |
This study aimed to determine the protein profiles and allergenic characteristics in
different body parts and genders of Scylla paramamosain. Raw and boiled extracts of a mix and six
individual body parts were prepared from the respective crab muscle tissues and then
analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE)
and immunoblotting using sera from 28 crab-allergic patients. The mix body part contains 24
protein fractions, while all individual body parts have almost similar protein profiles with 16 to
21 protein bands between 15 to
>250 kDa. The bands above 50 kDa were recognized as heat-sensitive proteins in the boiled extracts.
Therefore, the boiled extracts have fewer protein bands than the raw extracts. Both heat-sensitive
and heat-resistant protein bands have capabilities to bind to IgE antibodies of the tested
sera, but with dissimilar binding patterns. A heat- sensitive allergen at 95 kDa and four
heat-resistant allergens at 50, 42, 38 and 36 kDa were identified as the major allergens with the
frequencies of IgE-binding at 54, 50, 60, 64 and 54%, respectively. Immunoblotting of selected sera
revealed various IgE- binding patterns against individual body parts. The female leg was
identified as the most allergenic part than the other crab parts based on the number of
IgE-binding bands, number of major allergens and IgE-binding band intensities. Only a few bands
particularly the 38 kDa retain the IgE-binding capabilities in all boiled extracts. Mass
spectrometry analysis identified the 38 kDa of S. paramamosain as tropomyosin, while the
42 kDa were identified as beta-actin. As a conclusion, this study indicated that all body parts
of S. paramamosain have numerous heat-sensitive and heat- resistant allergenic proteins,
contributing to variable allergenicity between the crab body parts. Five protein bands at
95, 50, 42, 38 and 36 kDa were identified as the major allergens of this crab. The
implication is, these findings are useful for the advancement of the IgE-based diagnosis and
management of crab-allergic patients.
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