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Identification of a serine protease inhibitor homologue in Bird's Nest by an integrated proteomics approach
Author(s) -
Ou Keli,
Seow Teck K.,
Liang Rosa C. M. Y.,
Lee Bee W.,
Goh Denise L. M.,
Chua Kaw Y.,
Chung Maxey C. M.
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200109)22:16<3589::aid-elps3589>3.0.co;2-j
Subject(s) - serine protease , allergen , proteomics , tandem mass spectrometry , proteome , protease , nest (protein structural motif) , proteases , chemistry , egg white , gel electrophoresis , mass spectrometry , biology , microbiology and biotechnology , biochemistry , chromatography , allergy , immunology , enzyme , gene
For centuries, the edible nests of Collocalia spp. (“Bird's Nests”) have been used as a Chinese delicacy that had been claimed to be an effective health‐giving tonic. However, clinical studies indicated that in Singapore, Bird's Nest is the most common cause of food‐induced anaphylaxis in children, which could lead to potentially life‐threatening allergenic reactions. The purpose of this study was to characterize the major allergens in Bird's Nest by using the combined technologies of two‐dimensional gel electrophoresis (2‐DE), immunochemistry, N‐ terminal protein sequencing, and mass spectrometry. Results from the immunostaining of the Western blots of the Bird's Nest 2‐DE separated proteins with the sera from allergic patients indicated the presence of a major allergen of 66 kDa. Initial searches of the matrix assisted laser desorption/ionization – time of flight – mass spectrometry (MALDI‐TOF‐MS) tryptic peptide masses of the allergen in the SWISS‐PROT and NCBI nonredundant databases revealed that this protein was novel. Based on the partial protein sequence information obtained from N‐ terminal microsequencing and nanoelectrospray‐tandem MS, the 66 kDa immunoreactive allergen was found to be homologous to ovoinhibitor, a Kazal‐type serine protease inhibitor, which is one of the dominant allergens found in chicken egg white.