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Residues K128‐Q175 of human interleukin‐6 are essential for its biological activity
Author(s) -
Liu Haitian,
Duan Jubao,
Wang Jiaxi,
Peng Shanyun,
Zou Minji
Publication year - 1997
Publication title -
iubmb life
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.132
H-Index - 113
eISSN - 1521-6551
pISSN - 1521-6543
DOI - 10.1080/15216549700203501
Subject(s) - recombinant dna , mutant , plasmid , complementary dna , microbiology and biotechnology , receptor , biological activity , interleukin 2 , biology , in vitro , chemistry , cell culture , mtt assay , biochemistry , gene , genetics
Internal deletion K128‐Q175 of the human interleukin‐6 (hIL‐6) has been generated at the cDNA level. With pBV220 as expressing vector, the recombinant pBV*‐DIL‐6 encoding the deletion mutant (12kD) of hIL‐6 has been constructed. The resulted recombinant plasmids were then used to transform E. coli strain HB101, and the expression in the PLPR promoter system, which is temperature‐regulatable, was achieved. After purification and renaturation, the biological activity of the expressed product, designated as DM120, was measured by MTT method in an IL‐6‐dependent cell line 7TD1. The results show that the amino acid residues of IL‐6 128 to 175 are crucial for IL‐6 activity. Receptor binding assay in vitro indicates that the entire region is not involved in forming the receptor binding surface.