A novel tumor necrosis factor-alpha mutant with significantly enhanced cytotoxicity and receptor binding affinity.
A novel tumor necrosis factor-alpha mutant (mutant M3), in which Ser and Tyr at positions 52 and 56 were substituted by Ile and Phe, respectively, along with deletion of 7 N-terminal amino acids, was prepared and its biological activities were investigated. The mutant exhibited a 14- to 24-fold increase in the cytotoxicity relative to the wild-type TNF on various cancer cell lines. The binding affinity of the mutant to TNF-R55 and TNF-R75 receptors was over 10-fold higher than that of the wild-type. TNF-alpha and the mutant show similar CD spectra in the far-UV region, indicating that the overall structure was not influenced by the mutations. The production of highly potent TNF-alpha mutant utilizing increase of hydrophobicity in the region 52-56 may provide a structural basis for a design of optimized TNF-alpha as a therapeutic purpose.
Shin
N K
NK
Protein Engineering Laboratory, Hanhyo Institute of Technology, Taejon, Republic of Korea.
Lee
I
I
Chang
S G
SG
Shin
H C
HC
D000818:Animals; D015703:Antigens, CD / Q000378:metabolism*; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D051379:Mice; D009154:Mutation; D009369:Neoplasms / Q000188:drug therapy* / Q000378:metabolism; D009374:Neoplasms, Experimental / Q000188:drug therapy / Q000378:metabolism; D011487:Protein Conformation; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism*; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D013329:Structure-Activity Relationship; D014407:Tumor Cells, Cultured; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*
D015703:Antigens, CD; D018124:Receptors, Tumor Necrosis Factor; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; D014409:Tumor Necrosis Factor-alpha