Human alpha 1-acid glycoprotein as a model protein for glycoanalysis in baculovirus expression vector system

• Published in: Journal of Asia-Pacific entomology Vol. 18; no. 2; pp. 303 - 309
• Main Authors: Morokuma, Daisuke, Xu, Jian, Mon, Hiroaki, Hirata, Kazuma, Hino, Masato, Kuboe, Shoko, Yamashita, Mami, Kusakabe, Takahiro, Lee, Jae Man
• Format: Journal Article
• Language: English
• Published: Elsevier B.V 01.06.2015; 한국응용곤충학회
• Subjects: Alpha 1-acid glycoprotein; Baculoviridae; Baculovirus expression vector system; bioactive properties; blood proteins; glycoproteins; glycosylation; hemolymph; humans; larvae; N-glycosylation; polyacrylamide gel electrophoresis; polysaccharides; Silkworm; silkworms; 농수해양학; Baculovirus expression vector system; N-glycosylation; Alpha 1-acid glycoprotein; Silkworm
• ISSN: 1226-8615; 1876-7990; 1876-7790
• DOI: 10.1016/j.aspen.2015.03.006

Glycosylation is an important post-translational modification that confers various biological activities, structural stability, and inter-molecular interactions to proteins. Baculovirus expression vector system (BEVS) is widely used to produce recombinant glycoproteins, which may not be suitable for clinical use due to differences in the N-linked glycan structure between insects and mammals. It is necessary to develop an appropriate model protein-base platform for glycoanalysis to engineer the insect-type N-glycosylation pathway into human type efficiently. In this study, we employed human plasma protein alpha 1-acid glycoprotein (α1AGP). It was highly secreted from cultured silkworm cells and larvae when using the BEVS and glycosylated with insect type N-linked glycans. Interestingly, when separated on SDS-PAGE, the purified recombinant α1AGP secreted into silkworm haemolymph generated six distinct products from three alternative translates, suggesting that α1AGP has variations for the recognition or choice of glycosylation sites. [Display omitted] •Recombinant human α1AGP was expressed and highly glycosylated in silkworm cells and larvae using BEVS.•The N-linked glycan structure of the recombinant human α1AGP was identified as the paucimannosidic type.•α1AGP is a good model glycoprotein for analyzing the N-glycosylation alterations in insect-BEVS.