title: Molecular determinants of the interaction between human high molecular weight kininogen and Candida albicans cell wall: Identification of kininogen-binding proteins on fungal cell wall and mapping the cell wall-binding regions on kininogen molecule.
abstract: An excessive production of vasoactive and proinflammatory bradykinin-related peptides, the kinins, is often involved in the human host defense against microbial infections. Recent studies have shown that a major fungal pathogen to humans, Candida albicans, can bind the proteinaceous kinin precursor, the high molecular weight kininogen (HK) and trigger the kinin-forming cascade on the cell surface. In this work, we preliminarily characterized a molecular mechanism underlying the HK adhesion to the fungal surface by (i) identification of major kininogen-binding constituents on the candidial cell wall and (ii) mapping the cell wall-binding regions on HK molecule. A major fraction of total fungal kininogen-binding capacity was assigned to β-1,3-glucanase-extractable cell wall proteins (CWP). By adsorption of CWP on HK-coupled agarose gel and mass spectrometric analysis of the eluted material, major putative HK receptors were identified, including Als3 adhesin and three glycolytic enzymes, i.e., enolase 1, phosphoglycerate mutase 1 and triosephosphate isomerase 1. Using monoclonal antibodies directed against selected parts of HK molecule and synthetic peptides with sequences matching selected HK fragments, we assigned the major fungal cell wall-binding ability to a short stretch of amino acids in the C-terminal part of domain 3 and a large continuous region involving the C-terminal part of domain 5 and N-terminal part of domain 6 (residues 479-564). The latter characteristics of HK binding to C. albicans surface differ from those reported for bacteria and host cells.
authors.affiliation: Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics, and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors.forename: Justyna
authors.initials: J
authors.lastname: Karkowska-Kuleta
authors.forename: Sylwia
authors.initials: S
authors.lastname: Kedracka-Krok
authors.forename: Maria
authors.initials: M
authors.lastname: Rapala-Kozik
authors.forename: Wojciech
authors.initials: W
authors.lastname: Kamysz
authors.forename: Sylwia
authors.initials: S
authors.lastname: Bielinska
authors.forename: Anna
authors.initials: A
authors.lastname: Karafova
authors.forename: Andrzej
authors.initials: A
authors.lastname: Kozik
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000378:metabolism; D001665:Binding Sites; D002176:Candida albicans / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D002473:Cell Wall / Q000187:drug effects / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000378:metabolism; D043326:Glucan 1,3-beta-Glucosidase / Q000494:pharmacology; D006801:Humans; D025301:Hyphae / Q000187:drug effects / Q000378:metabolism; D019679:Kininogen, High-Molecular-Weight / Q000378:metabolism*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000378:metabolism; D010736:Phosphoglycerate Mutase / Q000378:metabolism; D010751:Phosphopyruvate Hydratase / Q000737:chemistry; D011485:Protein Binding; D025941:Protein Interaction Mapping / Q000379:methods*; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000378:metabolism; D014305:Triose-Phosphate Isomerase / Q000378:metabolism
chemical_list: C113620:ALS3 protein, Candida albicans; D000911:Antibodies, Monoclonal; D005656:Fungal Proteins; D019679:Kininogen, High-Molecular-Weight; D010446:Peptide Fragments; D011956:Receptors, Cell Surface; D043326:Glucan 1,3-beta-Glucosidase; D010751:Phosphopyruvate Hydratase; D014305:Triose-Phosphate Isomerase; D010736:Phosphoglycerate Mutase