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 .
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Amino Acid Sequence
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Chromatography, Gel
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Crystallization
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Crystallography, X-Ray
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Gene Expression*
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Humans
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Molecular Sequence Data
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Protein Binding
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Receptors, Cell Surface / metabolism
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Receptors, Urokinase Plasminogen Activator
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Recombinant Proteins / chemistry
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Recombinant Proteins / genetics
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Recombinant Proteins / isolation & purification
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Recombinant Proteins / metabolism
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Solubility
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Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
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Urokinase-Type Plasminogen Activator / chemistry*
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Urokinase-Type Plasminogen Activator / genetics
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Urokinase-Type Plasminogen Activator / isolation & purification
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Urokinase-Type Plasminogen Activator / metabolism*
Substances
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PLAUR protein, human
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Receptors, Cell Surface
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Receptors, Urokinase Plasminogen Activator
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Recombinant Proteins
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Urokinase-Type Plasminogen Activator