title: Tetranectin-binding site on plasminogen kringle 4 involves the lysine-binding pocket and at least one additional amino acid residue.
abstract: Kringle domains are found in a number of proteins where they govern protein-protein interactions. These interactions are often sensitive to lysine and lysine analogues, and the kringle-lysine interaction has been used as a model system for investigating kringle-protein interactions. In this study, we analyze the interaction of wild-type and six single-residue mutants of recombinant plasminogen kringle 4 expressed in Escherichia coli with the recombinant C-type lectin domain of tetranectin and trans-aminomethyl-cyclohexanoic acid (t-AMCHA) using isothermal titration calorimetry. We find that all amino acid residues of plasminogen kringle 4 found to be involved in t-AMCHA binding are also involved in binding tetranectin. Notably, one amino acid residue of plasminogen kringle 4, Arg 32, not involved in binding t-AMCHA, is critical for binding tetranectin. We also find that Asp 57 and Asp 55 of plasminogen kringle 4, which both were found to interact with the low molecular weight ligand with an almost identical geometry in the crystal of the complex, are not of equal functional importance in t-AMCHA binding. Mutating Asp 57 to an Asn totally eliminates binding, whereas the Asp 55 to Asn, like the Arg 71 to Gln mutation, was found only to decrease affinity.
authors.affiliation: Department of Molecular and Structural Biology, University of Aarhus, Denmark.
authors.forename: J H
authors.initials: JH
authors.lastname: Graversen
authors.forename: B W
authors.initials: BW
authors.lastname: Sigurskjold
authors.forename: H C
authors.initials: HC
authors.lastname: Thøgersen
authors.forename: M
authors.initials: M
authors.lastname: Etzerodt
mesh_terms: D001665:Binding Sites; D001798:Blood Proteins / Q000378:metabolism*; D002151:Calorimetry; D018082:Kringles; D037181:Lectins, C-Type; D008239:Lysine / Q000378:metabolism*; D016297:Mutagenesis, Site-Directed; D010958:Plasminogen / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D001798:Blood Proteins; D037181:Lectins, C-Type; C048648:tetranectin; D010958:Plasminogen; D008239:Lysine