title: Large-scale preparation of the delta10 form of staphylokinase by in vitro processing of recombinant staphylokinase with purified human plasminogen.
abstract: The authors have developed a rapid and convenient method for purification of a low molecular weight form (delta 10) of the bacterial plasminogen activator, staphylokinase. Recombinant staphylokinase is expressed in Escherichia coli, with an amino terminal extension that facilitated purification by immobilized metal-affinity chromatography. Purified staphylokinase is treated with human plasminogen, and the resulting truncated form is purified using a combination of immobilized metal affinity chromatography and hydrophobic interaction chromatography. Purified protein is characterized by amino terminal sequencing and in vitro plasminogen activation assay.
authors.affiliation: Center for Macromolecular Crystallography, University of Alabama at Birmingham 35294-0005, USA.
authors.forename: D
authors.initials: D
authors.lastname: Chattopadhyay
authors.forename: J E
authors.initials: JE
authors.lastname: Stewart
authors.forename: L J
authors.initials: LJ
authors.lastname: DeLucas
mesh_terms: D000595:Amino Acid Sequence; D004926:Escherichia coli / Q000235:genetics; D006801:Humans; D066298:In Vitro Techniques; D008666:Metalloendopeptidases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D008970:Molecular Weight; D010958:Plasminogen / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics
chemical_list: D011994:Recombinant Proteins; D010958:Plasminogen; D008666:Metalloendopeptidases; C022653:auR protein, Staphylococcus aureus