Protein expression and preliminary crystallographic analysis of amino-terminal fragment of urokinase-type plasminogen activator.
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 .
Zhao
GengXiang
G
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.
Yuan
Cai
C
Bian
ChuanBing
C
Hou
Xiaomin
X
Shi
XiaoLi
X
Ye
XiaoMing
X
Huang
ZiXiang
Z
Huang
MingDong
M
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*
C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D011994:Recombinant Proteins; D014568:Urokinase-Type Plasminogen Activator