Secretion of glycosylated human erythropoietin from yeast directed by the α-factor leader region

• Published in: Gene Vol. 79; no. 1; pp. 167 - 180
• Main Authors: Elliott, Steve, Giffin, James, Suggs, Sidney, Lau, Edward P., Banks, Allen R.
• Format: Journal Article
• Language: English
• Published: Lausanne Elsevier B.V 30.06.1989; Amsterdam Elsevier; New York, NY
• Subjects: Animals; Biological and medical sciences; Biological Transport; Biotechnology; DNA, Fungal - genetics; Endopeptidases - metabolism; erythropoietin; Erythropoietin - genetics; Erythropoietin - metabolism; Fundamental and applied biological sciences. Psychology; Gene Amplification; genes; Genes, Synthetic; Genetic engineering; Genetic technics; Glycosylation; Heterologous secretion; hormone; Humans; Immunoblotting; Mating Factor; Methods. Procedures. Technologies; Peptides - genetics; Peptides - metabolism; Plasmids; Promoter Regions, Genetic; Protein Processing, Post-Translational; Protein Sorting Signals - metabolism; proteolytic processing; recombinant DNA; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; regulated expression; Saccharomyces cerevisiae; Saccharomyces cerevisiae - genetics; Saccharomyces cerevisiae - metabolism; EPO; nt; SG-CAA; EndoH; YPD; bp; 2 μ; hormone; Ap; IFN; SD-CAA; KEX2; recombinant DNA; oligo; aa; PBS; KEX2, kex2; SDS; proteolytic processing; regulated expression; PAGE; Heterologous secretion; ER; SCE; LB; Saccharomyces cerevisiae; Human; Intracellular transport; Heterologous system; Yeast; Molecular processing; Leader sequence; Erythropoietin; Secretion; Glycosylation; Gene expression; Glycoprotein hormone; Biological activity; Fungi; Immunoblotting assay; Ascomycetes; Thallophyta
• ISSN: 0378-1119; 1879-0038
• DOI: 10.1016/0378-1119(89)90102-9

The pre-pro α-factor leader region of the yeast MFα 1 gene was used to direct the secretion of the human glycoprotein, erythropoietin (EPO), into the culture medium. An examination of the role of expression level on secretion of biologically active EPO indicated that there are several rate-limiting steps. These include processing of the α-factor-EPO precursor protein by the KEX2-encoded endoproteinase and transport of the protein through the secretory pathway. The rate-limiting steps for transport were early in the secretory pathway, probably from the endoplasmic reticulum to the Golgi apparatus.