Dataset Information

Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile) and the Effect of Cooking on Their Allergenicity.

ABSTRACT:

Objectives

To identify the major allergenic proteins of clam (Paphia textile) and to investigate the effect of different cooking methods on the allergenicity of these identified proteins.

Methods

Clam protein extracts were separated by denaturing polyacrylamide gel electrophoresis. IgE reactive proteins were then analyzed by immunoblotting with sera from patients with positive skin prick tests (SPT) to the raw clam extract. Mass spectrometry was used to identify the major allergenic proteins of this clam.

Results

Raw extract showed 12 protein bands (18-150 kDa). In contrast, fewer protein bands were seen in the boiled extract; those ranging from 40 to 150 kDa were denatured. The protein profiles were similarly altered by frying or roasting. The immunoblots of raw and boiled extracts yielded 10 and 2 IgE-binding proteins, respectively. The fried and roasted extracts showed only a single IgE-binding protein at 37 kDa. Mass spectrometry analysis of the 37 and 42 kDa major allergens indicated that these spots were tropomyosin and actin, respectively.

Conclusion

The two major allergens of Paphia textile were identified as the thermostable tropomyosin and a new thermolabile allergen actin.

SUBMITTER: Mohamad Yadzir ZH

PROVIDER: S-EPMC4564582 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

ACCESS DATA

Publications

Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile) and the Effect of Cooking on Their Allergenicity.

Mohamad Yadzir Zailatul Hani ZH Misnan Rosmilah R Bakhtiar Faizal F Abdullah Noormalin N Murad Shahnaz S

BioMed research international 20150827

<h4>Objectives</h4>To identify the major allergenic proteins of clam (Paphia textile) and to investigate the effect of different cooking methods on the allergenicity of these identified proteins.<h4>Methods</h4>Clam protein extracts were separated by denaturing polyacrylamide gel electrophoresis. IgE reactive proteins were then analyzed by immunoblotting with sera from patients with positive skin prick tests (SPT) to the raw clam extract. Mass spectrometry was used to identify the major allergen ...[more]

PMID: 26413512

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Project description:BACKGROUND:Many types of shellfish including oysters are sometime cooked before ingestion and it has been demonstrated that cooking may affect the allergenicity of food. Therefore, the aim of our present study is to identify major and minor allergens of tropical oyster (Crassostrea belcheri) and to investigate the effect of different cooking processing on the allergenicity of this oyster. METHODS:Raw, boiled, fried and roasted extracts of oyster were prepared. Protein profiles were analysed using sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). Major and minor allergens and allergenicity patterns of all extracts were then determined by immunoblotting with sera from patients with positive skin prick tests (SPT) to the raw oyster extract. Mass-spectrometry was used to identify the major allergenic proteins of this oyster. RESULTS:SDS-PAGE of the raw extract showed 15 protein bands (20-180 kDa). In contrast, smaller numbers of protein bands were demonstrated in the boiled extract, those ranging between 40-42 and 55-150 kDa were denatured, whereas the protein profiles were altered to a similar degree by frying or roasting. The 37 kDa proteins had the highest frequency of IgE-binding (95 %), thus identified as the major allergen of this tropical oyster. Other minor IgE-binding proteins were observed at various molecular weights. Immunoblot of raw extract yielded 11 IgE-binding proteins. The cooked extracts showed only a single IgE-binding protein at 37 kDa. Mass spectrometry analysis of the 37 kDa major allergen identified this spot as tropomyosin. CONCLUSIONS:Cooked extracts produce lower IgE-binding than raw extract, which suggest that thermal treatment can be used as a tool in attempting to reduce oyster allergenicity by reducing the number of IgE-reactive bands. The degree of allergenicity of this oyster was demonstrated in the order raw > boiled > fried ≈ roasted. A heat-resistent 37 kDa protein, corresponding to tropomyosin, was identified as the major allergen of this tropical oyster.

Project description:BackgroundMost aquatic biota's reproductive biology and life cycle are essential to the sustainable management and development of coastal ecosystems and aquaculture. The bivalve Paphia textile (Gmelin 1791), also known as Paratapes textilis, has an economic value in Indo-Pacific waters, including the Red Sea and the Suez Canal lakes, the Egyptian coasts. However, P. textile suffers from extensive fishing and exploitation.AimThe present work aims to study the Paphia textile's reproductive cycle on the Egyptian coasts of the Red Sea for the first time. It helps to manage and develop the coastal ecosystems and aquaculture.MethodologySamples were collected monthly from two saline lakes in the Suez Gulf from December 2019 to November 2020. As part of the comprehensive research study, sex ratio, condition index, sexuality, histological analysis of gonads, shell size, and gonad index were used to investigate the reproductive cycle.ResultsThe results reveal a male-biased sex ratio, possibly due to anthropogenic stressors. The Paphia textile is dioecious. No hermaphrodite cases were observed in the studied specimens. The condition index in winter and spring indicates periods dominated by mature individuals. Five reproductive maturity stages were assigned for both P. textile males and females. Due to the simultaneous development of several developmental stages monthly throughout the sampling year, warm water may be responsible for non-sequential gametogenic cycles. As measured environmental parameters correlate with maturity stages, temperature, salinity, and chlorophyll a play important role in gonad growth. The size at first sexual maturity at which 50% of the Paphia textile population reached maturity ranged from 28.60 to 31.50 mm for females, and between 31.70 and 34.10 mm for males. As the gonad index increases during the ripe stages, this index decreases during the resting, spawning, and spent phases.ConclusionsThe findings suggest the most suitable temperature for aquaculture spawning is between 20 °C and 30 °C in subtropical waters. Fishing should generally be prohibited at sizes less than 28.60 mm for better management and sustainability of this valuable aquatic resource on the Egyptian coasts of the Red Sea.

Dataset Information

Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile) and the Effect of Cooking on Their Allergenicity.

Objectives

Methods

Results

Conclusion

Publications

Tropomyosin and Actin Identified as Major Allergens of the Carpet Clam (Paphia textile) and the Effect of Cooking on Their Allergenicity.

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