title: The impact of N-glycosylation on conformation and stability of immunoglobulin Y from egg yolk.
abstract: Immunoglobulin Y (IgY) is a new therapeutic antibody, and its applications in industry are very broad. To provide insight into the effects of N-glycosylation on IgY, its conformation and stability were studied. In this research, IgY was extracted from egg yolk and then digested by peptide-N4-(N-acetyl-beta-glucosaminyl) asparagine-amidase. SDS-PAGE and infrared absorption spectrum showed that carbohydrates were distinctly reduced after enzymolysis. The circular dichroism spectrum indicated that the IgY molecule became more flexible and disordered after removal of N-glycan. The fluorescence intensity revealed that Trp residues were buried in a more hydrophobic environment after disposal of N-glycan. Storage stability decreased with the removal of oligosaccharide chains based on size-exclusion chromatography analysis. Deglycosylated IgY exhibited less resistance to guanidine hydrochloride-induced unfolding. After deglycosylation, IgY was more sensitive to pepsin. Therefore, N-glycosylation played an important role in the maintenance of the structure and stability of IgY.
authors.affiliation: National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
authors.forename: Long
authors.initials: L
authors.lastname: Sheng
authors.affiliation: National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
authors.forename: Zhenjiao
authors.initials: Z
authors.lastname: He
authors.affiliation: National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
authors.forename: Jiahui
authors.initials: J
authors.lastname: Chen
authors.affiliation: National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
authors.forename: Yaofa
authors.initials: Y
authors.lastname: Liu
authors.affiliation: National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China.
authors.forename: Meihu
authors.initials: M
authors.lastname: Ma
authors.affiliation: National Research and Development Center for Egg Processing, College of Food Science and Technology, Huazhong Agricultural University, Wuhan, Hubei 430070, PR China. Electronic address: caizhaoxia@mail.hzau.edu.cn.
authors.forename: Zhaoxia
authors.initials: Z
authors.lastname: Cai
mesh_terms: D000818:Animals; D004530:Egg Yolk / Q000737:chemistry*; D006031:Glycosylation; D019791:Guanidine / Q000494:pharmacology; D007136:Immunoglobulins / Q000737:chemistry* / Q000378:metabolism*; D009584:Nitrogen / Q000378:metabolism*; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / Q000378:metabolism; D011487:Protein Conformation; D011489:Protein Denaturation / Q000187:drug effects; D055550:Protein Stability; D059748:Proteolysis
chemical_list: C016512:IgY; D007136:Immunoglobulins; D043524:Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase; D019791:Guanidine; D009584:Nitrogen
keywords: Conformation; Immunoglobulin Y; N-glycosylation