Human interleukin 1 beta 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-1 beta (IL-1 beta) 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-1 beta, 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-1 beta without a signal peptide produced the predicted 17.5-kDa nonglycosylated form. These results suggest that fusion to a heterologous leader sequence allowed IL-1 beta to be translocated across the membrane of the endoplasmic reticulum and to be transported and secreted by the exocytotic pathway.
Pecceu
F
F
Unité Technologie des Cellules Animales Recombinées, Sanofi Elf Bio Recherches, Labège, France.
Dousset
P
P
Shire
D
D
Cavrois
E
E
Marchese
E
E
Ferrara
P
P
Kaghad
M
M
Dumont
X
X
Lupker
J
J
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D003001:Cloning, Molecular; D006224:Cricetinae; D015870:Gene Expression; D006031:Glycosylation; D013006:Growth Hormone / Q000235:genetics*; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D011233:Precipitin Tests; D021382:Protein Sorting Signals / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014415:Tunicamycin / Q000494:pharmacology
D007375:Interleukin-1; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D014415:Tunicamycin; D013006:Growth Hormone