title: Release of biologically active kinin peptides, Met-Lys-bradykinin and Leu-Met-Lys-bradykinin from human kininogens by two major secreted aspartic proteases of Candida parapsilosis.
abstract: In terms of infection incidence, the yeast Candida parapsilosis is the second after Candida albicans as causative agent of candidiases in humans. The major virulence factors of C. parapsilosis are secreted aspartic proteases (SAPPs) which help the pathogen to disseminate, acquire nutrients and dysregulate the mechanisms of innate immunity of the host. In the current work we characterized the action of two major extracellular proteases of C. parapsilosis, SAPP1 and SAPP2, on human kininogens, proteinaceous precursors of vasoactive and proinflammatory bradykinin-related peptides, collectively called the kinins. The kininogens, preferably the form with lower molecular mass, were effectively cleaved by SAPPs, with the release of two uncommon kinins, Met-Lys-bradykinin and Leu-Met-Lys-bradykinin. While optimal at acidic pH (4-5), the kinin release yield was only 2-3-fold lower at neutral pH. These peptides were able to interact with cellular kinin receptors of B2 subtype and to stimulate the human endothelial cells HMEC-1 to increased secretion of proinflammatory interleukins (ILs), IL-1β and IL-6. The analysis of the stability of SAPP-generated kinins in plasma suggested that they are biologically equivalent to bradykinin, the best agonist of B2 receptor subtype and can be quickly converted to des-Arg(9)-bradykinin, the agonist of inflammation-inducible B1 receptors.
authors: Department of Analytical Biochemistry, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Gronostajowa 7, 30-387 Krakow, Poland.
authors: Grazyna
authors: G
authors: Bras
authors: Oliwia
authors: O
authors: Bochenska
authors: Maria
authors: M
authors: Rapala-Kozik
authors: Ibeth
authors: I
authors: Guevara-Lora
authors: Alexander
authors: A
authors: Faussner
authors: Wojciech
authors: W
authors: Kamysz
authors: Andrzej
authors: A
authors: Kozik
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000378:metabolism*; D057055:Aspartic Acid Proteases; D001920:Bradykinin / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism; D002175:Candida / Q000201:enzymology / Q000472:pathogenicity; D002177:Candidiasis / Q000201:enzymology* / Q000382:microbiology / Q000473:pathology; D042783:Endothelial Cells; D005656:Fungal Proteins / Q000378:metabolism*; D006801:Humans; D007704:Kininogens / Q000737:chemistry / Q000378:metabolism*; D009842:Oligopeptides; D010455:Peptides / Q000737:chemistry / Q000378:metabolism
chemical_list: D005656:Fungal Proteins; D007704:Kininogens; D009842:Oligopeptides; D010455:Peptides; C006791:bradykinin potentiating factors; C018103:bradykinin, Met-Lys-; D057055:Aspartic Acid Proteases; D016282:Aspartic Acid Endopeptidases; C079801:SAPP1 protein, Candida parapsilosis; D001920:Bradykinin
keywords: 3,3′,5,5′-tetramethylbenzidine; ACE; ACN; B1 receptor; B1R; B2 receptor; B2R; BK; BSA; Bradykinin receptors; CBB; Candida parapsilosis; Candidiasis; Coomassie Brilliant Blue dye; DL-2-mercaptomethyl-3-guanidinoethylthiopropionic acid; DMEM; DTT; Dulbecco's modified Eagle's medium; EBM; EGF; ELISA; ESI; ESI-MS/MS; ESI-tandem mass spectrometry; ETD; FBS; FPLC; HC; HK; HKD4; HPLC; HRP; HRP-conjugated streptavidin; IL-1β; IL-6; ILs; IT; Interleukins; Kallikrein–kinin system; LC–MS; LK; LPS; MGTA; MS; NAC; PBS; RP-HPLC; SA-HRP; SAPs; SDS-PAGE; Secreted aspartic proteases; TFA; TMB; UHPLC; YCB; YPD; acetonitrile; angiotensin-converting enzyme; bovine serum albumin; bradykinin; dithiotreitol; domain 4 of high molecular mass kininogen; electron transfer dissociation; electrospray ionization; endothelial basal medium; enzyme-linked immunosorbent assay; epithelial growth factor; fast protein liquid chromatography; fetal bovine serum; high molecular mass kininogen; high-performance liquid chromatography; horseradish peroxidase; hydrocortizon; interleukin-1β; interleukin-6; interleukins; ion-trap; lipopolysaccharide; liquid chromatography–mass spectrometry; low molecular mass kininogen; mass spectrometry; phosphate-buffered saline; reversed-phase HPLC; secreted aspartic proteases; sodium dodecyl sulfate polyacrylamide gel electrophoresis; trifluoroacetic acid; ultra HPLC; yeast carbone base; yeast peptone dextrose medium; “non-albicans” Candida species