A molecular and in silico characterisation of Hev b 4 a glycosylated latex allergen


Citation

A. Malik, . and S. Ahmad, . and Siti Arija M. Arif, . and E. Sunderasan, . (2007) A molecular and in silico characterisation of Hev b 4 a glycosylated latex allergen. [Proceedings Paper]

Abstract

Hev b 4 lecithinase homologue is a highly glycosylated allergenic latex protein with seven attached N-glycan structures comprising of oligomannose and complex type N-glycans. Treatment with a mixture of N-glycosidase A and N-glycosidase F resulted in lowering of the band corresponding to Hev b 4 on SDS-gel from 53-55 kDa to circa 40 kDa this being comparable to the 38.53 kDa mass predicted by its cDNA. In Western-immunoblots the enzymatically deglycosylated Hev b 4 showed negligible binding to sera immunoglobulin E lgE from latex allergic patients; however recognition to anti-Hev b 4 rabbit polyclonal antibody lgG was retained. The results of the Western immunoblots indicated that IgE binding ability of Hev b 4 is essentially determined by its N-glycan moiety presumably by the complex type N-glycans. The complete coverage of the amino acid sequence and the N-glycan structures enabled the construction of a homology based 3D model of He v b 4. Six out of seven N-glycans were already incorporated in the preliminary 3D model of Hev b 4; when completed the model will open the way to the delineation of IgE binding to the glyco-allergen.


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Abstract

Hev b 4 lecithinase homologue is a highly glycosylated allergenic latex protein with seven attached N-glycan structures comprising of oligomannose and complex type N-glycans. Treatment with a mixture of N-glycosidase A and N-glycosidase F resulted in lowering of the band corresponding to Hev b 4 on SDS-gel from 53-55 kDa to circa 40 kDa this being comparable to the 38.53 kDa mass predicted by its cDNA. In Western-immunoblots the enzymatically deglycosylated Hev b 4 showed negligible binding to sera immunoglobulin E lgE from latex allergic patients; however recognition to anti-Hev b 4 rabbit polyclonal antibody lgG was retained. The results of the Western immunoblots indicated that IgE binding ability of Hev b 4 is essentially determined by its N-glycan moiety presumably by the complex type N-glycans. The complete coverage of the amino acid sequence and the N-glycan structures enabled the construction of a homology based 3D model of He v b 4. Six out of seven N-glycans were already incorporated in the preliminary 3D model of Hev b 4; when completed the model will open the way to the delineation of IgE binding to the glyco-allergen.

Additional Metadata

[error in script]
Item Type: Proceedings Paper
AGROVOC Term: LATEX
AGROVOC Term: HEVEA BRASILIENSIS
AGROVOC Term: MOLECULAR BIOLOGY
AGROVOC Term: DIALYSIS
AGROVOC Term: ELECTROPHORESIS
AGROVOC Term: ANALYTICAL METHODS
AGROVOC Term: AMINO ACIDS
AGROVOC Term: IMMUNOBLOTTING
AGROVOC Term: IMMUNOGLOBULINS
AGROVOC Term: BIOTECHNOLOGY
Geographical Term: MALAYSIA
Depositing User: Ms. Norfaezah Khomsan
Last Modified: 24 Apr 2025 05:27
URI: http://webagris.upm.edu.my/id/eprint/15700

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