title: Expression, purification and characterization of yeast protein disulfide isomerase produced by a recombinant baculovirus-mediated silkworm, Bombyx mori, pupae expression system.
abstract: Protein disulfide isomerase (PDI) is a multifunctional polypeptide presents in the endoplasmic reticulum of the cell. Silkworm (Bombyx mori) pupae were used as hosts to produce recombinant PDI (rPDI). The concentration-dependent chaperone activity of rPDI was evidenced by the inhibition of the aggregation of rhodanese. Approximately 297 microg rPDI was purified from a single silkworm pupa. Results of rPDI treated with endoglycosidase H and N-glycanase, PNGase F, indicate that non-N-glycosylated rPDI (occupying 90%) and N-glycosylated rPDI are expressed in the silkworm expression system. The difference in glycosylation between silkworm pupae and yeast is discussed.
authors.affiliation: Institute of Environmental Science for Human Life, Department of Food and Nutritional Sciences, Ochanomizu University, 2-1-1 Ohtsuka, Bunkyo-ku, Tokyo 112-8610, Japan. mimihuahuaer@yahoo.co.jp
authors.forename: Liyun
authors.initials: L
authors.lastname: Wang
authors.forename: Yuri
authors.initials: Y
authors.lastname: Shimizu
authors.forename: Takemitsu
authors.initials: T
authors.lastname: Mizunaga
authors.forename: Shogo
authors.initials: S
authors.lastname: Matsumoto
authors.forename: Yuzuru
authors.initials: Y
authors.lastname: Otsuka
mesh_terms: D000818:Animals; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D012831:Bombyx / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D006031:Glycosylation; D019704:Protein Disulfide-Isomerases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D011679:Pupa / Q000235:genetics; D011994:Recombinant Proteins / Q000302:isolation & purification / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology* / Q000235:genetics
chemical_list: D005656:Fungal Proteins; D011994:Recombinant Proteins; D019704:Protein Disulfide-Isomerases