Mutagenesis of human granulocyte colony stimulating factor |
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Authors: | T Kuga Y Komatsu M Yamasaki S Sekine H Miyaji T Nishi M Sato Y Yokoo M Asano M Okabe |
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Affiliation: | Tokyo Research Laboratories, Kyowa Hakko Kogyo Co. Ltd., Japan. |
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Abstract: | To define the structure-function relationship, we have made a number of mutants of human granulocyte colony-stimulating factor (hG-CSF) by in vitro mutagenesis. The results indicate that most of the mutations located in the internal and C-terminal regions of the molecule abolished the activity, whereas the mutants without N-terminal 4, 5, 7, or 11 amino acids retained the activity. N-terminal amino acids were also altered by cassette mutagenesis using a synthetic oligonucleotide mixture. Among them, KW2228, in which Thr-1, Leu-3, Gly-4, Pro-5 and Cys-17 were respectively substituted with Ala, Thr, Tyr, Arg and Ser, showed more potent granulopoietic activity than that of intact hG-CSF both in vitro and in vivo. |
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