Abstract
The pre-pro alpha-factor leader region of the yeast MF alpha 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 alpha-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.
MeSH terms
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Animals
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Biological Transport
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DNA, Fungal / genetics
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Endopeptidases / metabolism
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Erythropoietin / genetics
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Erythropoietin / metabolism*
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Gene Amplification
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Genes, Synthetic
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Glycosylation
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Humans
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Immunoblotting
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Mating Factor
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Peptides / genetics*
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Peptides / metabolism
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Plasmids
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Promoter Regions, Genetic
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Protein Processing, Post-Translational
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Protein Sorting Signals / metabolism*
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Saccharomyces cerevisiae / genetics*
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Saccharomyces cerevisiae / metabolism
Substances
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DNA, Fungal
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Peptides
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Protein Sorting Signals
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Recombinant Fusion Proteins
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Erythropoietin
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Mating Factor
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Endopeptidases