Human interleukin 1β fused to the human growth hormone signal peptide is N-glycosylated and secreted by Chinese hamster ovary cells
A hybrid gene consisting of the sequences coding for the signal peptide of human growth hormone and the mature form of interleukin-1β (IL-1β) was chemically synthesized. This sequence was inserted into a eukaryotic expression vector and introduced into Chinese hamster ovary cells. The resulting stab...
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Veröffentlicht in: | Gene 1991-01, Vol.97 (2), p.253-258 |
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Sprache: | eng |
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Zusammenfassung: | A hybrid gene consisting of the sequences coding for the signal peptide of human growth hormone and the mature form of interleukin-1β (IL-1β) was chemically synthesized. This sequence was inserted into a eukaryotic expression vector and introduced into Chinese hamster ovary cells. The resulting stably transformed cell lines produced large amounts of recombinant IL-1β, which was secreted into the culture medium mainly as a 22-kDa form. Expression in the presence of tunicamycin, an inhibitor of
N-glycosylation, led to the complete disappearance of the 22-kDa form and the appearance of a new form of 17.5 kDa, indicating that the hybrid protein had been both processed and
N-glycosylated. However, transformed cells producing mature IL-1β without a signal peptide produced the predicted 17.5-kDa nonglycosylated form. These results suggest that fusion to a heterologous leader sequence allowed IL-1β to be translocated across the membrane of the endoplasmic reticulum and to be transported and secreted by the exocytotic pathway. |
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ISSN: | 0378-1119 1879-0038 |
DOI: | 10.1016/0378-1119(91)90059-K |