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

2015 ◽  
Vol 2015 ◽  
pp. 1-6 ◽  
Author(s):  
Zailatul Hani Mohamad Yadzir ◽  
Rosmilah Misnan ◽  
Faizal Bakhtiar ◽  
Noormalin Abdullah ◽  
Shahnaz Murad
Keyword(s):  
Mass Spectrometry ◽  
Polyacrylamide Gel Electrophoresis ◽  
Mass Spectrometry Analysis ◽  
Cooking Methods ◽  
Positive Skin ◽  
Spectrometry Analysis ◽  
Ige Binding ◽  
Allergenic Proteins ◽  
Skin Prick Tests ◽  
Major Allergens

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 ofPaphia textilewere identified as the thermostable tropomyosin and a new thermolabile allergen actin.

scholarly journals CHARACTERIZATION OF MAJOR ALLERGENS OF MACROBRACHIUM ROSENBERGII (GIANT RIVER PRAWN) BY ALLERGENOMIC APPROACH AND THEIR THERMOSTABILITY PROPERTIES

2018 ◽  
Vol 81 (1) ◽  
Author(s):  
Rosmilah Misnan ◽  
Komathi Sockalingam ◽  
Zailatul Hani Mohamad Yadzir ◽  
Noormalin Abdullah ◽  
Faizal Bakhtiar ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Polyacrylamide Gel Electrophoresis ◽  
Arginine Kinase ◽  
Sodium Dodecyl ◽  
Macrobrachium Rosenbergii ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Heat Treated ◽  
Major Allergens ◽  
Prawn Allergy

Prawn allergy is a common cause of allergic reactions in countries where crustacean is a famous seafood. Macrobrachium rosenbergii (giant river prawn) is declared as a local major food allergens, causes severe symptoms like asthma and anaphylactic shock. Therefore, the goal concerning this research is to identify the allergenicity of heat treated of M. rosenbergii. Prawn extracts have been prepared using fresh raw prawn and three heat-treated prawn flesh (boiled, steamed, fried), then afterward analyzed by the usage of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) in conformity with their protein profiles. Allergenic proteins were identified by means of immunoblotting by the use of sera from 30 prawn-allergic patients. The identities of selective major allergens were then determined by mass-spectrometry analysis. The raw prawn has 27 protein fractions between 6 to 207 kDa, while the heat-treated prawns have reduced number of protein bands. Six prominent bands at 72, 65, 48, 38, 36, and 30 kDa were considered as the major allergens of raw extract. Meanwhile, after heat remedies applied, most of the IgE-binding bands were disappeared except for three major allergens at 38, 36 or 30 kDa which were still remain to be seen, suggesting as highly thermostable allergens. Among heat-treated prawns, steamed and boiled elicited more allergenic bands, contrast to fried prawn extract. Mass spectrometry analysis identified two selected major allergens at 36 and 48 kDa as tropomyosin and arginine kinase, respectively. As a conclusion, this study indicated that both thermostable and thermolabile proteins from M. rosenbergii were allergenic. Tropomyosin and arginine kinase were identified as the most important major allergens. Thus, the knowledge on thermostability of prawn allergens are crucial for improving diagnosis and management of prawn allergic patients in this county. 

scholarly journals PgTI, the First Bioactive Protein Isolated from the Cactus Pilosocereus gounellei, is a Trypsin Inhibitor with Antimicrobial Activity

2019 ◽  
pp. 1-11
Author(s):  
Cláudio Alberto Alves Da Rocha Filho ◽  
Poliana Karla Amorim ◽  
Thâmarah de Albuquerque Lima ◽  
Pollyanna Michelle da Silva ◽  
Maiara Celine de Moura ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Flow Cytometry ◽  
Antimicrobial Activity ◽  
Trypsin Inhibitor ◽  
Polyacrylamide Gel Electrophoresis ◽  
Mass Spectrometry Analysis ◽  
Minimal Bactericidal Concentration ◽  
Trypsin Inhibitor Activity ◽  
Minimal Inhibitory Concentrations ◽  
Spectrometry Analysis

Aims: This work aimed to isolate, characterize and evaluate the antimicrobial activity of a trypsin inhibitor (PgTI) from the stem of Pilosocereus gounellei. Place and Duration of Study: Departamento de Bioquímica, Universidade Federal de Pernambuco between March 2013 and October 2018. Instituto de Bioquímica Médica Leopoldo de Meis, Universidade Federal do Rio de Janeiro between June and July 2018. Methodology: PgTI was isolated from P. gounellei stem extract by gel filtration and ion exchange chromatographies. The inhibitor was characterized by isoelectric focusing, polyacrylamide gel electrophoresis, tryptic digestion followed by mass spectrometry analysis and for stability towards heating. Antibacterial and antifungal activities were investigated through broth microdilution assays. Viability of the microbial cells was also evaluated by flow cytometry analysis using thiazol orange and propidium iodide. Results: PgTI appeared as a single polypeptide band of 37.1 kDa and isoelectric point (pI) 5.88. The inhibition constant (Ki) for bovine trypsin was 14 nM and mass spectrometry analysis of PgTI did not reveal similarities with other plant proteins. Trypsin inhibitor activity was stable at temperatures up to 50ºC. PgTI inhibited growth of Gram-positive and Gram-negative bacteria (minimal inhibitory concentrations (MIC) from 7.5 to150 µg/mL) with bactericidal activity only against Escherichia coli (minimal bactericidal concentration: 75.0 µg/mL). PgTI also inhibited the growth of Candida krusei (MIC of 60 µg/mL). Flow cytometry confirmed that PgTI did not affect the viability of E. coli and C. krusei cells at the MIC. Conclusion: This is the first report on a bioactive protein purified from P. gounellei, which provides biotechnological value to this cactus.

Identification of IgE-binding proteins of Thunnus tonggol (tongkol) by mass spectrometry analysis

2007 ◽  
Vol &NA; ◽  
pp. S93
Author(s):  
Rosmilah Misnan ◽  
Shahnaz Murad ◽  
Meinir Jones ◽  
Graham Taylor ◽  
Dinah Rahman ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Binding Proteins ◽  
Mass Spectrometry Analysis ◽  
Spectrometry Analysis ◽  
Ige Binding

scholarly journals Structural Characterization of an Oligosaccharide Made by Neisseria sicca

2009 ◽  
Vol 191 (10) ◽  
pp. 3311-3320 ◽  
Author(s):  
Ellen T. O'Connor ◽  
Hui Zhou ◽  
Kevin Bullock ◽  
Karen V. Swanson ◽  
J. McLeod Griffiss ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Structural Characterization ◽  
Lectin Binding ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Mass Spectrometry Analysis ◽  
Ionization Mass ◽  
Spectrometry Analysis ◽  
Neisseria Sicca

ABSTRACT Neisseria sicca 4320 expresses two carbohydrate-containing components with sodium dodecyl sulfate-polyacrylamide gel electrophoresis mobilities that resemble those of lipooligosaccharide and lipopolysaccharide. Using matrix-assisted laser desorption ionization—time of flight and electrospray ionization mass spectrometry, we characterized a disaccharide carbohydrate repeating unit expressed by this strain. Gas chromatography identified the sugars composing the unit as rhamnose and N-acetyl-d-glucosamine. Glycosidase digestion confirmed the identity of the nonreducing terminal sugar of the disaccharide and established its β-anomeric configuration. Mass spectrometry analysis and lectin binding were used to verify the linkages within the disaccharide repeat. The results revealed that the disaccharide repeat is [-4) β-l-rhamnose (1-3) β-N-acetyl-d-glucosamine (1-] with an N-acetyl-d-glucosamine nonreducing terminus. This work is the first structural characterization of a molecule that possesses rhamnose in the genus Neisseria.

IDENTIFICATION OF TROPOMYOSIN AS THE MAJOR ALLERGEN OF BLACK TIGER PRAWN (PENAEUS MONODON) BY AN ALLERGENOMIC APPROACH

2015 ◽  
Vol 77 (25) ◽  
Author(s):  
Rosmilah Misnan ◽  
Noor Asyikin Kamarazaman ◽  
Zailatul Hani Mohd Yadzir ◽  
Noormalin Abdullah ◽  
Mohd Faizal Bakhtiar ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Penaeus Monodon ◽  
Major Allergen ◽  
Mass Spectrometry Analysis ◽  
Asia Pacific ◽  
Spectrometry Analysis ◽  
Ige Binding ◽  
Black Tiger Prawn ◽  
Tiger Prawn ◽  
Prawn Allergy

Shellfish has been recognized as one of the leading causes of food allergy in both adults and children in Asia Pacific region. In Malaysia, black tiger prawn (Penaeus monodon) is among the most widely consumed species. Our previous studies have successfully identified several major allergens including a thermostable protein of 36 kDa. Thus the aim of this study was to identify the 36 kDa major allergen by an allergenomic approach. Protein extracts of raw prown were prepared and resolved by 2-dimensional electrophoresis (2-DE). Immunoblotting was then performed using sera from patients with prawn allergy. Selected spot from 2-DE was then excised, digested and analyzed by mass spectrometry. The 2-DE profile of the extract revealed approximately 100 protein spots between pH of ~4 to 10 and the size range between (<10 to 250 kDa). The 2-DE immunoblotting has detected numerous IgE-binding spots at 36 kDa. Mass spectrometry analysis of the major IgE-binding spot (spot 1a) has identified the 36 kDa spot as tropomyosin. Our findings indicated that tropomyosin play a major role in allergic reaction to black tiger prawn among local patients with prawn allergy, and should be included in diagnostics and therapeutic strategies of this allergy.

Characterization of Major Allergens of Local Mud Crab (Scylla serrata)

2015 ◽  
Vol 12 (1) ◽  
pp. 1 ◽  
Author(s):  
Rosmilah Misnan ◽  
Nurul Izzah Abdul Rahman ◽  
Zailatul Hani Mohd Yadzir ◽  
Noormalin Abdullah ◽  
Mohd Faizal Bakhtiar ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Arginine Kinase ◽  
Mass Spectrometry Analysis ◽  
Scylla Serrata ◽  
Mud Crab ◽  
Meat Allergy ◽  
Proteomic Approach ◽  
The World ◽  
Major Allergens ◽  
Crab Meat

Crab meat is widely consumed in several countries around the world. However, when consumed, crab meats are frequent cause of allergic reactions throughout the world. Scylla serrata is among the most common mud crab in Malaysia. In a previous study two major allergens of mud crab at 36 and 41 kDa was identified. Thus, the aim of this study is to further identify these major allergens by a proteomic approach. Protein extract was prepared and resolved by 2-dimensional electrophoresis (2-DE). Immunoblotting was then performed using reactive sera from patients with crab allergy. Major allergenic spots were then excised from the 2-DE gel and analysed by mass spectrometry. The 2-DE profile of the extract revealed approximately >100 protein spots between pH of 4.00 to 8.00. Mass spectrometry analysis has identified the 36 and 41 kDa proteins as tropomyosin and arginine kinase, respectively. Our findings indicated that tropomyosin and arginine kinase play a major role in allergic reaction to mud crab meat among local patients with crab meat allergy, and should be included in diagnostics and therapeutic strategies of this allergy.

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