The hsp150 delta-carrier confers secretion competence to the rat nerve growth factor receptor ectodomain in Saccharomyces cerevisiae
| Autor: | M, Simonen, H, Vihinen, E, Jämsä, U, Arumäe, N, Kalkkinen, M, Makarow |
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| Rok vydání: | 1996 |
| Předmět: |
Glycosylation
Saccharomyces cerevisiae Proteins Base Sequence Glycoside Hydrolases beta-Fructofuranosidase Recombinant Fusion Proteins Molecular Sequence Data Biological Transport Receptors Nerve Growth Factor Saccharomyces cerevisiae Protein Sorting Signals Endoplasmic Reticulum Receptor Nerve Growth Factor Peptide Fragments Rats Tumor Cells Cultured Animals Humans Disulfides Nerve Growth Factors Melanoma Heat-Shock Proteins Glycoproteins |
| Zdroj: | Yeast (Chichester, England). 12(5) |
| ISSN: | 0749-503X |
| Popis: | When the extracellular domain of rat low-affinity nerve growth factor receptor (NGFRe) was synthesized in Saccharomyces cerevisiae with the signal peptide of invertase, NGFRe was translocated to the endoplasmic reticulum (ER) and retained there. However, when NGFRe was fused to the C-terminus of the hsp150 delta-carrier, the hsp150 delta-NGFRe fusion protein was efficiently secreted to the growth medium with no apparent retention in the ER. The NGFRe portion was disulphide-bonded and its single N-glycosylation site was occupied. The hsp150 delta-carrier is an N-terminal signal peptide-containing fragment of a yeast secretory glycoprotein. Hsp150 delta-NGFRe, harvested from the culture medium, inhibited the cross-linking of [125I]NGF to authentic NGFR on the surface of human melanoma cells. Moreover, [125I]NGF could be chemically cross-linked to secretory hsp150 delta-NGFRe, suggesting that the NGFRe portion had adopted a ligand-binding conformation. However, inhibition of the cross-linking by unlabelled NGF was less effective than in the case of the authentic receptor. The hsp150 delta-carrier may have potential in the production of mammalian proteins, which require elaborate folding and disulphide formation in the ER. |
| Databáze: | OpenAIRE |
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