Efficient production of biologically active porcine interleukin-18 by coexpression with porcine caspase-1 using a baculovirus expression system.
We previously reported that the precursor form of porcine interleukin-18 (IL-18) expressed by the baculovirus system was able to be secreted efficiently into the supernatant of insect cells, whereas only small amounts of mature IL-18 were secreted from insect cells. As insect cells do not normally have the IL-1beta converting enzyme (caspase-1), which is required for processing of the precursor IL-18 into the mature IL-18, we recently cloned porcine caspase-1 cDNA. In this study, we constructed a recombinant baculovirus containing the cDNA encoding porcine caspase-1 and showed that the coexpression of caspase-1 and the precursor IL-18 enabled insect cells to secrete mature IL-18 into the culture supernatant efficiently. Moreover, inhibition of caspase-1 activity by its specific inhibitor prevented the processing of precursor IL-18 into the mature form. These results indicated that the processing and secretion of precursor IL-18 into the mature form in insect cells were enhanced by the artificial introduction of caspase-1 activity for cleavage.
Muneta
Y
Y
Institute of Animal Health, Ibaraki 305-0856, Japan. ymuneta@niah.affrc.go.jp
Inumaru
S
S
Shimoji
Y
Y
Mori
Y
Y
D000818:Animals; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D020170:Caspase 1 / Q000235:genetics* / Q000378:metabolism; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D020382:Interleukin-18 / Q000096:biosynthesis* / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism; D018411:Spodoptera; D013552:Swine; D014162:Transfection
D017931:DNA Primers; D020382:Interleukin-18; D011994:Recombinant Proteins; D020170:Caspase 1