Expression and activity of a recombinant chimeric protein composed of pokeweed antiviral protein and of human interleukin-2.
The pokeweed antiviral protein (PAP) has already been used to chemically construct immunotoxins. Here we tested the recombinant approach for the production of PAP-containing cytotoxic fusion-proteins. A cDNA encoding a mutated PAP (PAP9), which is expressed at high levels in bacteria, was fused to human interleukin-2 (IL-2) cDNA. The resulting PAP9-IL-2 protein was as active as the free PAP9 in inhibiting an eukaryotic cell-free translation system. Only the chimeric protein desaminated the 28S rRNA and inhibited translation of the CTLL-2 cell line which expresses the IL-2 receptor. These results show that PAP is a suitable toxin for the production of recombinant immunotoxins.
Dore
J M
JM
Diaclone, Besançon, France.
Gras
E
E
Wijdenes
J
J
D000818:Animals; D002460:Cell Line; D006801:Humans; D000922:Immunotoxins; D007376:Interleukin-2 / Q000235:genetics / Q000494:pharmacology*; D051379:Mice; D009699:N-Glycosyl Hydrolases; D010940:Plant Proteins / Q000235:genetics / Q000494:pharmacology*; D014176:Protein Biosynthesis / Q000187:drug effects; D012339:RNA, Ribosomal, 28S / Q000378:metabolism; D015375:Receptors, Interleukin-2 / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000494:pharmacology*; D054789:Ribosome Inactivating Proteins, Type 1
D000922:Immunotoxins; D007376:Interleukin-2; D010940:Plant Proteins; D012339:RNA, Ribosomal, 28S; D015375:Receptors, Interleukin-2; D011993:Recombinant Fusion Proteins; D054789:Ribosome Inactivating Proteins, Type 1; D009699:N-Glycosyl Hydrolases; C024275:pokeweed antiviral protein