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Evidence that the conformation of unliganded human plasminogen is maintained via an intramolecular interaction between the lysine-binding site of kringle 5 and the N-terminal peptide

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Original languageEnglish
Pages (from-to)99-105
Number of pages7
JournalBiochemical Journal
Publication statusPublished - 1 Jul 1998


Human Glu-plasminogen adopts at least three conformations that provide a means for regulating the specificity of its activation in vivo. It has been proposed previously that the closed (cr) conformation of human Glu-plasminogen is maintained through physical interaction of the kringle 5 domain and a lysine residue within the N-terminal peptide (NTP), To examine this hypothesis, site-directed mutagenesis was used to generate variant proteins containing substitutions either for aspartic acid residues within the anionic centre of the kringle 5 domain or for conserved lysine residues within the NTP, Size-exclusion HPLC and rates of plasminogen activation by urokinase-type plasminogen activator were used to determine the conformational states of these variants. Variants with substitutions within the kringle 5 lysine-binding site demonstrated extended conformations, as did variants with alanine substitutions for Lys(50) and Lys(62), In contrast, molecules in which NTP residues Lys(20) Or Lys(33) were replaced were shown to adopt closed conformations. We conclude that the lysine-binding site of kringle 5 is involved in maintaining the closed conformation of human Glu-plasminogen via an interaction with the NTP, probably through Lys(50) and/or Lys(62) These conclusions advance the current model for the initial stages of fibrinolysis during which fibrin is thought to compete with the NTP for the kringle 5 lysine-binding site.

    Research areas

  • Amino Acid Substitution, Aminocaproic Acid, Animals, Binding Sites, CHO Cells, Chromatography, High Pressure Liquid, Cricetinae, Enzyme Activation, Glutamic Acid, Humans, Kringles, Ligands, Lysine, Molecular Weight, Mutagenesis, Site-Directed, Plasminogen, Protein Conformation, Recombinant Proteins, Urokinase-Type Plasminogen Activator

ID: 24447547