The substitution of cysteine 17 of recombinant human G-CSF with alanine greatly enhanced its stability. |
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| Authors: | M Ishikawa H Iijima R Satake-Ishikawa H Tsumura A Iwamatsu T Kadoya Y Shimada H Fukamachi K Kobayashi S Matsuki |
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| Institution: | Pharmaceutical Laboratory, Kirin Brewery Co., Ltd., Gunma, Japan. |
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| Abstract: | 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, Ile, 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. Among these 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. |
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