title: The substitution of cysteine 17 of recombinant human G-CSF with alanine greatly enhanced its stability.
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.
authors.affiliation: Pharmaceutical Laboratory, Kirin Brewery Co., Ltd., Gunma, Japan.
authors.forename: M
authors.initials: M
authors.lastname: Ishikawa
authors.forename: H
authors.initials: H
authors.lastname: Iijima
authors.forename: R
authors.initials: R
authors.lastname: Satake-Ishikawa
authors.forename: H
authors.initials: H
authors.lastname: Tsumura
authors.forename: A
authors.initials: A
authors.lastname: Iwamatsu
authors.forename: T
authors.initials: T
authors.lastname: Kadoya
authors.forename: Y
authors.initials: Y
authors.lastname: Shimada
authors.forename: H
authors.initials: H
authors.lastname: Fukamachi
authors.forename: K
authors.initials: K
authors.lastname: Kobayashi
authors.forename: S
authors.initials: S
authors.lastname: Matsuki
mesh_terms: D000409:Alanine / Q000737:chemistry* / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells; D003545:Cysteine / Q000737:chemistry* / Q000235:genetics; D005260:Female; D016179:Granulocyte Colony-Stimulating Factor / Q000737:chemistry* / Q000235:genetics / Q000502:physiology; D006358:Hot Temperature; D006801:Humans; D066298:In Vitro Techniques; D051379:Mice; D008807:Mice, Inbred BALB C; D016297:Mutagenesis, Site-Directed / Q000502:physiology; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics; D013329:Structure-Activity Relationship
chemical_list: D011994:Recombinant Proteins; D016179:Granulocyte Colony-Stimulating Factor; D003545:Cysteine; D000409:Alanine