title: Overexpression of human calnexin in yeast improves measles surface glycoprotein solubility.
abstract: The limitations of high-level expression of virus surface proteins in yeast are not well understood. The inefficiency of yeast to produce active human virus surface glycoproteins, as well as other mammalian glycoproteins, is usually explained by the inefficient folding of the glycoprotein into its characteristic and functional three-dimensional structure from a random coil. The endoplasmic reticulum (ER) is a highly versatile protein factory that is equipped with chaperones and folding enzymes essential for protein folding. To improve folding and solubility of viral surface glycoprotein, the genes encoding human ER resident chaperones calnexin, calreticulin, immunoglobin binding protein (BiP), protein disulfide isomerase (PDI) and foldase (ERp57) were coexpressed together with hemagglutinin gene from measles virus in the yeast Saccharomyces cerevisiae. The effect of coexpressing chaperones on the total yield of measles virus hemagglutinin (MeH) as well as the intracellular fate of the glycoprotein was determined. Our results demonstrated that coexpression of human calnexin noticeably enhanced the quantity of the soluble glycosylated form of MeH in yeast. The coexpression of human calreticulin-, PDI-, ERp57- and BiP-encoding genes did not improve the quality of recombinant MeH.
authors.affiliation: Institute of Biotechnology, Vilnius University, Lithuania.
authors.forename: Evaldas
authors.initials: E
authors.lastname: Ciplys
authors.forename: Kęstutis
authors.initials: K
authors.lastname: Sasnauskas
authors.forename: Rimantas
authors.initials: R
authors.lastname: Slibinskas
mesh_terms: D037281:Calnexin / Q000096:biosynthesis* / Q000235:genetics*; D015870:Gene Expression; D006389:Hemagglutinins, Viral / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D012995:Solubility
chemical_list: D006389:Hemagglutinins, Viral; D011994:Recombinant Proteins; C032397:hemagglutinin protein G, measles virus; D037281:Calnexin