Abstract
Bovine follicle-stimulating hormone (bFSH), a pituitary gonadotropin, is a heterodimer hormone that consists of a common alpha-subunit non-covalently associated with the hormone-specific beta-subunit. Unfortunately, expression levels of recombinant bFSH or its subunits are invariably low. We report here the secretory expression of biologically active bFSHalpha and bFSHbeta subunit in the methylotrophic yeast Hansenula polymorpha. A slightly higher level of expression of recombinant bFSH subunits was achieved by using the Saccharomyces cerevisiae-derived calnexin (ScCne1) as a chaperone in engineered H. polymorpha strains. The preliminary data also suggested that bFSH subunits expressed in H. polymorpha appeared to be less-glycosylated. This isoform had been shown to be 80% increase in in vivo bioactivity compared with the hyperglycosylated Pichia pastoris-derived recombinant bFSHalpha/beta. More sophisticated applications of bFSH would profit from the assembled less-glycosylated heterodimer.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Animals
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Calnexin / genetics
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Calnexin / metabolism
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Cattle
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Chromatography, Affinity
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Codon / metabolism
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Follicle Stimulating Hormone, beta Subunit / biosynthesis*
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Follicle Stimulating Hormone, beta Subunit / chemistry
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Follicle Stimulating Hormone, beta Subunit / genetics
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Follicle Stimulating Hormone, beta Subunit / metabolism*
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Glycoprotein Hormones, alpha Subunit / biosynthesis*
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Glycoprotein Hormones, alpha Subunit / chemistry
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Glycoprotein Hormones, alpha Subunit / genetics
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Glycoprotein Hormones, alpha Subunit / metabolism
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Glycosylation
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Histidine / chemistry
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Histidine / genetics
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Oligopeptides / chemistry
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Oligopeptides / genetics
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Pichia / genetics*
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Recombinant Fusion Proteins / biosynthesis*
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Recombinant Fusion Proteins / chemistry
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Recombinant Fusion Proteins / genetics
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Recombinant Fusion Proteins / metabolism
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Saccharomyces cerevisiae Proteins / genetics
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Saccharomyces cerevisiae Proteins / metabolism
Substances
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CNE1 protein, S cerevisiae
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Codon
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Follicle Stimulating Hormone, beta Subunit
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Glycoprotein Hormones, alpha Subunit
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His-His-His-His-His-His
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Oligopeptides
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Recombinant Fusion Proteins
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Saccharomyces cerevisiae Proteins
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Calnexin
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Histidine