The Substitution of Cysteine 17 of Recombinant HumanG-CSF with Alanine Greatly Enhanced its Stability

Cell Structure and Function

Hiromi Fukamachi²  Keiko Kobayashi²  Hiroshi Iijima¹  Rika Satake-Ishikawa²  Katsuhiko Asano²  Haruhiko Tsumura²  Masaharu Ishikawa²  Akihiro Iwamatsu¹  Shigeru Matsuki²  Toshihiko Kadoya²  Yoshihiro Shimada² 

[1] Central Laboratories for Key Technology, Kirin Brewery Co., Ltd.;Pharmaceutical Laboratory, Kirin Brewery Co., Ltd.

关键词: cysteine;    granulocyte-colony stimulating factor;    site-directed mutagenesis;    thermostability;   

DOI  :  10.1247/csf.17.61

学科分类：分子生物学，细胞生物学和基因

来源: Japan Society for Cell Biology

【 摘 要 】

References(25)Cited-By(11)Human recombinant granulocyte-colony stimulating factor (rhG-CSF) has one free cysteine at position 17 and has two disulfide bridges (Cys36-Cys42 and Cys64-Cys74). The Cys17 of rhG-CSF was substituted with Gly, Ala, Ser, He, Tyr, Arg, and Pro, or deleted using site-directed mutagenesis in order to improve its thermostability. With the exception of Pro17-rhG-CSF, all mutant proteins retained biological activity which promotes the growth of mouse bone marrow cells in vitro. Amongthese mutant proteins, Ala17-rhG-CSF had more than 5 times higher stability than rhG-CSF. But Ser17-rhG-CSF had almost same stability as rhG-CSF and other mutant proteins had only lower stability.

【 授权许可】

Unknown