Synthesis and maturation of recombinant human tumor necrosis factor in eukaryotic systems.
The biosynthesis of human tumor necrosis factor (hTNF) was studied. The amino-terminal extension of the hTNF precursor (26 kDa polypeptide) was not cleaved off in a cell-free system supplemented with dog pancreas microsomes. Correct maturation of pre-hTNF was nevertheless not restricted to the macrophage system: in the medium of a TNF-producing, transformed CHO cell line, a (weak) approximately 20 kDa, an approximately 18.5 kDa (doublet) and a 17 kDa TNF polypeptide, the latter corresponding to mature hTNF, were revealed by specific immunoprecipitation. Similar results were obtained with Xenopus laevis oocytes, injected with hTNF mRNA, except that the 20 kDa band was lacking. The results are discussed in relation to the secretion mechanism of hTNF.
D000818:Animals; D002474:Cell-Free System; D006224:Cricetinae; D004274:DNA, Recombinant / Q000378:metabolism*; D004285:Dogs; D005260:Female; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D007163:Immunosorbent Techniques; D008861:Microsomes / Q000378:metabolism; D009865:Oocytes / Q000378:metabolism; D010053:Ovary / Q000378:metabolism; D010179:Pancreas / Q000648:ultrastructure; D014176:Protein Biosynthesis; D012315:RNA Caps / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D011994:Recombinant Proteins; D012156:Reticulocytes / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha; D014982:Xenopus laevis
D004274:DNA, Recombinant; D006023:Glycoproteins; D012315:RNA Caps; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha