A molecular and in silico characterisation of Hev b 4, a glycosylated latex allergen
2007
A. Malik, Jamia Millia Islamia Univ, New Delhi (India) | S. Ahmad, National Institute of Biomedical Innovation, Ibaraki-city, Osaka (Japan) | Siti Arija M. Arif | E. Sunderasan, Rubber Research Institute of Malaysia, Malaysia Rubber Board, P. O. Box 10150, 50908 Kuala Lumpur (Malaysia)
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.
Mostrar más [+] Menos [-]Palabras clave de AGROVOC
Información bibliográfica
Este registro bibliográfico ha sido proporcionado por Universiti Putra Malaysia