title: Protein expression and preliminary crystallographic analysis of amino-terminal fragment of urokinase-type plasminogen activator.
abstract: The amino-terminal fragment (ATF, Ser1-Glu143) of urokinase-type plasminogen activator (uPA) is responsible for some important functions of uPA, such as receptor binding and chemotactic activity. To dissect the function and structure-activity relationship of ATF, recombinant human ATF was expressed in Pichia pastoris system at a yield of about 30 mg/L. The recombinant ATF was captured by a cation exchange column, further purified up to 99% purity by a gel filtration column, and characterized in terms of its receptor binding capability. The purified ATF was then crystallized by the method of sitting-drop vapor diffusion with magnesium sulfate as the precipitating agent at 298 K. The crystals belong to space group P1 with unit cell dimensions of a=47.5A, b=64.7A, c=65.4A, alpha=71.6 degrees , beta=92.1 degrees , gamma=84.0 degrees .
authors.affiliation: State Key Laboratory of Structural Biology, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, 155 Yang Qiao Xi Lu, Fujian 350002, China.
authors.forename: GengXiang
authors.initials: G
authors.lastname: Zhao
authors.forename: Cai
authors.initials: C
authors.lastname: Yuan
authors.forename: ChuanBing
authors.initials: C
authors.lastname: Bian
authors.forename: Xiaomin
authors.initials: X
authors.lastname: Hou
authors.forename: XiaoLi
authors.initials: X
authors.lastname: Shi
authors.forename: XiaoMing
authors.initials: X
authors.lastname: Ye
authors.forename: ZiXiang
authors.initials: Z
authors.lastname: Huang
authors.forename: MingDong
authors.initials: M
authors.lastname: Huang
mesh_terms: D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D003460:Crystallization; D018360:Crystallography, X-Ray; D015870:Gene Expression; D006801:Humans; D008969:Molecular Sequence Data; D011485:Protein Binding; D011956:Receptors, Cell Surface / Q000378:metabolism; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014568:Urokinase-Type Plasminogen Activator / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*
chemical_list: C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator