Characterization of the Carbohydrate Moiety in a Partly Purified Allergen Preparation from the Mould Cladosporium herbarum and Its Possible Importance for Allergenic Activity as Tested by RAST-Inhibition

1984 ◽  
Vol 75 (2) ◽  
pp. 149-156 ◽  
Author(s):  
Marit Swärd-Nordmo ◽  
Berit Smestad Paulsen ◽  
Jens Kristian Wold ◽  
Øivind Grimmer
Keyword(s):  
Carbohydrate Moiety ◽  
Cladosporium Herbarum ◽  
Allergenic Activity

scholarly journals Expression and characterization of recombinant Par j 1 and Par j 2 resembling the allergenic epitopes of Parietaria judaica pollen

2019 ◽  
Vol 9 (1) ◽  
Author(s):  
Yulia Dorofeeva ◽  
Paolo Colombo ◽  
Miguel Blanca ◽  
Adriano Mari ◽  
Roman Khanferyan ◽  
...  
Keyword(s):  
Insect Cell ◽  
Binding Capacity ◽  
Gel Filtration ◽  
Mediterranean Area ◽  
Lipid Transfer ◽  
Allergen Specific Immunotherapy ◽  
Allergenic Activity ◽  
Allergen Sources ◽  
Monomeric Proteins

Abstract The weed wall pellitory, Parietaria judaica, is one the most important pollen allergen sources in the Mediterranean area causing severe symptoms of hay fever and asthma in allergic patients. We report the expression of the major Parietaria allergens, Par j 1 and Par j 2 which belong to the family of lipid transfer proteins, in insect cells. According to circular dichroism analysis and gel filtration, the purified allergens represented folded and monomeric proteins. Insect cell-expressed, folded Par j 2 exhibited higher IgE binding capacity and more than 100-fold higher allergenic activity than unfolded Escherichia coli-expressed Par j 2 as demonstrated by IgE ELISA and basophil activation testing. IgE ELISA inhibition assays showed that Par j 1 and Par j 2, contain genuine and cross-reactive IgE epitopes. IgG antibodies induced by immunization with Par j 2 inhibited binding of allergic patients IgE to Par j 1 only partially. IgE inhibition experiments demonstrated that insect cell-expressed Par j 1 and Par j 2 together resembled the majority of allergenic epitopes of the Parietaria allergome and therefore both should be used for molecular diagnosis and the design of vaccines for allergen-specific immunotherapy of Parietaria allergy.

scholarly journals Molecular Characterization of Allergens of Cladosporium herbarum and Alternaria alternans

1995 ◽  
Vol 107 (1-3) ◽  
pp. 458-459 ◽  
Author(s):  
Michael Breitenbach ◽  
Gernot Achatz ◽  
Hannes Oberkofler ◽  
Birgit Simon ◽  
Andrea Unger ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Cladosporium Herbarum

Further structural studies of the carbohydrate moiety of the allergen Ag-54 (Cla h II) from the mould Cladosporium herbarum

1991 ◽  
Vol 214 (2) ◽  
pp. 267-279 ◽  
Author(s):  
Marit Swärd-Nordmo ◽  
Berit Smestad Paulsen ◽  
Jens Kristian Wold ◽  
Thomas Wehler ◽  
Per-Erik Jansson
Keyword(s):  
Carbohydrate Moiety ◽  
Structural Studies ◽  
Cladosporium Herbarum

scholarly journals Isolation and partial characterization of two antigenic glycoproteins from rye-grass (Lolium perenne) pollen

1981 ◽  
Vol 197 (3) ◽  
pp. 695-706 ◽  
Author(s):  
B J Howlett ◽  
A E Clarke
Keyword(s):  
Lolium Perenne ◽  
Concanavalin A ◽  
Sodium Dodecyl Sulphate ◽  
Lectin Binding ◽  
Sodium Dodecyl ◽  
Binding Assays ◽  
Pea Lectin ◽  
Rye Grass ◽  
Allergenic Activity

Two glycoproteins have been purified from a buffer extract of rye-grass (Lolium perenne) pollen. Both migrated as single bands on sodium dodecyl sulphate/polyacrylamide gels. Glycoprotein 1 (0.8 mg/g of pollen) had a apparent mol.wt. of 33 000 and contained 95% protein and 5% carbohydrate. The monosaccharides glucose, galactose, mannose, arabinose and N-acetylglucosamine were present in the proportions 3:3:1:2:1. Glycoprotein 2 (0.4 mg/g of pollen) had an apparent mol. wt. of 68000 and contained 88% protein and 12% carbohydrate. The monosaccharides glucose, galactose, mannose, fucose, xylose, arabinose and N-acetylglucosamine were present in the proportions 4:7:13:5:8:6:6. This glycoprotein bound concanavalin A and Lotus tetragonolobus (asparagus pea) lectin. Radioallergosorbent (RAST) inhibition tests showed that Glycoprotein 1 is an effective allergen, whereas Glycoprotein 2 has less allergenic activity. A method for performing both lectin-binding assays and RAST inhibition tests using microtitre trays is described.

scholarly journals Hazelnut (Corylus avellana) vicilin Cor a 11: molecular characterization of a glycoprotein and its allergenic activity

2004 ◽  
Vol 383 (2) ◽  
pp. 327-334 ◽  
Author(s):  
Iris LAUER ◽  
Kay FOETISCH ◽  
Daniel KOLARICH ◽  
Barbara K. BALLMER-WEBER ◽  
Amedeo CONTI ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Corylus Avellana ◽  
Cd Spectroscopy ◽  
Mediator Release ◽  
Food Allergies ◽  
Ige Reactivity ◽  
Allergenic Activity ◽  
The Impact

In Europe, hazelnuts (Corylus avellana) are a frequent cause of food allergies. Several important hazelnut allergens have been previously identified and characterized. Specific N-glycans are known to induce strong IgE responses of uncertain clinical relevance, but so far the allergenic potential of glycoproteins from hazelnut has not been investigated. The aim of the study was the molecular characterization of the glycosylated vicilin Cor a 11 from hazelnut and the analysis of its allergenic activity. Although MALDI–TOF (matrix-assisted laser-desorption ionization–time-of-flight) MS showed that one of two potential glycosylation sites of Cor a 11 was glycosylated, CD spectroscopy indicated that recombinant and natural Cor a 11 share similar secondary structures. Thus to analyse the impact of the glycan residues of Cor a 11 on IgE binding, the allergenic activity of natural glycosylated Cor a 11 and recombinant Cor a 11 was compared. In addition, the IgE sensitization pattern to recombinant Cor a 11, Cor a 1, Cor a 2 and Cor a 8 of 65 hazelnut allergic patients was determined in vitro. The prevalence of IgE reactivity to hazelnut vicilin Cor a 11 was below 50%. Basophil histamine-release assays were used to determine the allergenic activity of both natural and recombinant Cor a 11 in comparison with Cor a 1, a birch (Betula verrucosa) pollen-related major hazelnut allergen. Both forms of Cor a 11 induced mediator release from basophils to a similar extent, indicating that the hazelnut allergic patients had cross-linking IgE antibodies binding to the protein backbone and not to carbohydrate structures. In comparison to Cor a 1, a 10000-fold higher concentration of Cor a 11 was required to induce similar basophil mediator release. In conclusion, the hazelnut vicilin Cor a 11 is a minor allergen both in regard to prevalence and allergenic potency, whereas its glycan does not contribute to its allergenic activity.

Characterization Of The Allergenic Activity Of Tropomyosin From Aedes Aegypti

2014 ◽  
Vol 133 (2) ◽  
pp. AB102
Author(s):  
Jose F. Cantillo ◽  
Leonardo Puerta ◽  
Enrique Fernandez-Caldas ◽  
Sylvie Lafosse-Marin
Keyword(s):  
Aedes Aegypti ◽  
Allergenic Activity

scholarly journals Characterization of the invertase from Pichia anomala

1995 ◽  
Vol 306 (1) ◽  
pp. 235-239 ◽  
Author(s):  
J Rodriguez ◽  
J A Perez ◽  
T Ruiz ◽  
L Rodriguez
Keyword(s):  
Carbon Source ◽  
Molecular Mass ◽  
Total Mass ◽  
Culture Medium ◽  
Thermal Sensitivity ◽  
Apparent Molecular Mass ◽  
Carbohydrate Moiety ◽  
Pichia Anomala

Synthesis of invertase (EC 3.2.1.26) in Pichia anomala is controlled by the carbon source in the culture medium. The enzyme was purified to homogeneity from P. anomala cells fully derepressed for invertase synthesis and shown to be a multimeric glycoprotein composed of identical subunits with an apparent molecular mass of 86.5 kDa. The carbohydrate moiety accounts for approx. 30% of the total mass of the molecule and consists of manno-oligosaccharides N-linked to the polypeptide. Most of the characteristics of the enzyme analysed in this study were similar to those previously reported for other yeast invertases, with the remarkable exception of its thermal sensitivity which appears after 15 min incubation at temperatures above 32 degrees C.

scholarly journals Purification and characterization of the invertase from Schizosaccharomyces pombe. A comparative analysis with the invertase from Saccharomyces cerevisiae

1990 ◽  
Vol 267 (3) ◽  
pp. 697-702 ◽  
Author(s):  
S Moreno ◽  
Y Sanchez ◽  
L Rodriguez
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Schizosaccharomyces Pombe ◽  
Molecular Mass ◽  
Total Mass ◽  
High Molecular Mass ◽  
Carbohydrate Moiety ◽  
Purification And Characterization ◽  
Protein Moiety ◽  
Protein Portion

Invertase (EC 3.2.1.26) was purified to homogeneity from exponentially growing cells of Schizosaccharomyces pombe fully de-repressed for synthesis of the enzyme, and was shown to be a high-molecular-mass glycoprotein that can be dissociated in the presence of 8 M-urea/1% SDS into identical subunits with an apparent molecular mass of 205 kDa. The carbohydrate moiety, accounting for 67% of the total mass, is composed of equimolar amounts of mannose and galactose. There is a small amount of glucosamine, which is probably involved in the linkage to the protein moiety, since the enzyme is sensitive to treatment with endoglycosidase H. The composition of the carbohydrate moiety resembles that found in higher-eukaryotic glycoproteins and differs from glycoproteins found in Saccharomyces cerevisiae. The protein portion of each subunit is a polypeptide of molecular mass 60 kDa, very similar to the invertase of Sacch. cerevisiae. Both proteins cross-react with antibodies raised against the protein fractions of the other, indicating that the two enzymes are similar.

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