Mutational analysis of human tumor necrosis factor-alpha.
To understand the structure-function relationship of human tumor necrosis factor-alpha (TNF-alpha), mutational analysis was carried out on the lower regions (regions 1-6) of the molecule. The muteins were prepared as a soluble form by using a chaperonin co-expression system and the cytotoxic activities of the purified muteins were evaluated on TNF-sensitive murine fibrosarcoma L929 cells. Three regions (regions 1, 2 & 4) were found where mutations significantly influenced the bioactivity. In region 1 (residues 1-10), the number of deleted residues and the positioning of positive charges are important to achieve a maximum activity and in region 4 (residues 84-88), introduction of charged residues in one of the positions 86-88 significantly increased the cytotoxic activity. On the other hand, any mutation introduced in region 2 (residues 37-41) had a deleterious effect. The present study provides a structural basis for the design of highly potent TNF-alpha as a therapeutic agent.
Shin
Hang-Cheol
HC
Department of Bioinformatics and Life Science, and Computer Aided Molecular Design Research Center, Soongsil University, Seoul, Korea, 156-743. hcshin@ssu.ac.kr
Cho
Kwang-Hwi
KH
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000818:Animals; D001665:Binding Sites; D005354:Fibrosarcoma / Q000188:drug therapy; D006801:Humans; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D013329:Structure-Activity Relationship; D018675:Toxicity Tests; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000737:chemistry* / Q000235:genetics* / Q000494:pharmacology
D014409:Tumor Necrosis Factor-alpha