Solution Structure of Human Plasminogen Kringle 3
[PLMN_HUMAN] Defects in PLG are the cause of plasminogen deficiency (PLGD) [MIM:217090]. PLGD is characterized by decreased serum plasminogen activity. Two forms of the disorder are distinguished: type 1 deficiency is additionally characterized by decreased plasminogen antigen levels and clinical symptoms, whereas type 2 deficiency, also known as dysplasminogenemia, is characterized by normal, or slightly reduced antigen levels, and absence of clinical manifestations. Plasminogen deficiency type 1 results in markedly impaired extracellular fibrinolysis and chronic mucosal pseudomembranous lesions due to subepithelial fibrin deposition and inflammation. The most common clinical manifestation of type 1 deficiency is ligneous conjunctivitis in which pseudomembranes formation on the palpebral surfaces of the eye progresses to white, yellow-white, or red thick masses with a wood-like consistency that replace the normal mucosa.       
[PLMN_HUMAN] Plasmin dissolves the fibrin of blood clots and acts as a proteolytic factor in a variety of other processes including embryonic development, tissue remodeling, tumor invasion, and inflammation. In ovulation, weakens the walls of the Graafian follicle. It activates the urokinase-type plasminogen activator, collagenases and several complement zymogens, such as C1 and C5. Cleavage of fibronectin and laminin leads to cell detachment and apoptosis. Also cleaves fibrin, thrombospondin and von Willebrand factor. Its role in tissue remodeling and tumor invasion may be modulated by CSPG4. Binds to cells. Angiostatin is an angiogenesis inhibitor that blocks neovascularization and growth of experimental primary and metastatic tumors in vivo.
Publication Abstract from PubMed
Human plasminogen kringle 3 (hPgn K3) domain contains most elements of the canonical lysine binding site (LBS) found in other Pgn kringles. However, it does not exhibit affinity for either lysine or structurally related zwitterionic ligands. It has been shown that lysine-binding activity can be engineered via a Lys57 --> Asp mutation [Burgin J and Schaller J, Cell. Mol. Life Sci. 55, 135, 1999]. Using a recombinant construct expressed in E. coli, the three-dimensional solution structure of hPgn K3 was determined via NMR spectroscopy [heavy atom averaged RMSD = 0.35 +/- 0.07 A (backbone) and 0.75 +/- 0.12 A (all)]. The 1H/15N heteronuclear single quantum correlated (HSQC) spectra for both wild-type K3 and mutated [r(K57D)K3] structures are essentially identical, implying that the two structures are effectively isomorphous. The affinity of r(K57D)K3 for the lysine analog trans-(aminomethyl)-cyclohexanecarboxylic acid (AMCHA) was investigated from ligand-induced NMR chemical shift perturbations, which enabled for mapping the binding site on the mutated domain surface. The equilibrium association constant, Ka, was determined to be ~ 5.23 +/- 0.03 mM-1. Homology modeling combined with in silico docking of lysine-like zwitterionic ligands via AutoDock 4.0 supports functionality of the engineered (K57D)K3 LBS, whose electrostatic focal centers are defined by the Arg36/Arg71 cationic and Asp55/Asp57 anionic pairs. Comparison of K3-type sequences from different vertebrates, including kringles from hedgehog apolipoprotein(a) [Apo-(a)] and Apo(a)-related [Arp] sequences, reveals that Lys57 is confined to the hPgn variant. Based on the likely phylogeny and ligand affinities of the homologous domains, it is suggested that the hPgn K3 is "aberrant" in that all other K3-type domains, including hedgehog Apo(a) and all Arp domains, except K3(1), are predicted to variously exhibit lysine-binding capability. In Arp K3(1) an Arg residue fills site 72 replacing the key Trp residue found in all other kringles, thus interfering with a requisite kringle - ligand hydrophobic interaction.
Human Plasminogen Kringle 3: Solution Structure, Functional Insights, Phylogenetic Landscape.,Christen MT, Frank P, Schaller J, Llinas M Biochemistry. 2010 Jul 9. PMID:20617841
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.