title: Molecular characterization, expression analysis, and bactericidal activity of the derivative peptides of TFPI-1 and TFPI-2 in half-smooth tongue sole, Cynoglossus semilaevis.
abstract: Tissue factor pathway inhibitors (TFPIs) are Kunitz-type serine protease inhibitors that reversibly regulate the blood coagulation induced by tissue factor. TFPI family contain two members, TFPI-1 and TFPI-2. Recent studies have shown TFPI-1 and TFPI-2 also play important roles in innate immunity, however, the potential function of teleost TFPI are very limited. In this study, we characterized two TFPI (CsTFPI-1 and CsTFPI-2) molecules from half-smooth tongue sole (Cynoglossus semilaevis), examined their tissue distributions and expression patterns under pathogens stimulation as well as investigated the antibacterial activity of the C-terminal peptides. Quantitative real time RT-PCR analysis showed that constitutive CsTFPI-1 expression occurred, in increasing order, in head kidney, intestine, brain, spleen, liver, skin, gills, heart, and muscle; CsTFPI-2 was expressed, in increasing order, in the gills, intestine, skin, head kidney, liver, brain, spleen, muscle, and heart. Under Vibrio anguillarum, Streptococcus agalactiae and fish megalocytivirus stimulation, both CsTFPI-1 and CsTFPI-2 expression increased significantly in a manner that depended on the pathogen, tissue type, and infection stage, which suggested CsTFPI-1 and CsTFPI-2 play important roles in anti-bacterial and anti-viral infection. Finally, C-terminal peptides of CsTFPI-1 and CsTFPI-2, were synthesized and proved to have antibacterial effect against Micrococcus luteus that were independent of host serum. Take together, these results indicate that CsTFPI-1 and CsTFPI-2 play important roles in antimicrobial immunity of this fish.
authors.affiliation: Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
authors.forename: Xin-Peng
authors.initials: XP
authors.lastname: Zhao
authors.affiliation: Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
authors.forename: Shu-Wen
authors.initials: SW
authors.lastname: He
authors.affiliation: Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
authors.forename: Bin
authors.initials: B
authors.lastname: Yue
authors.affiliation: Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China.
authors.forename: Guang-Hua
authors.initials: GH
authors.lastname: Wang
authors.affiliation: Marine Science and Engineering College, Qingdao Agricultural University, Qingdao, 266109, China. Electronic address: lissazhang06@163.com.
authors.forename: Min
authors.initials: M
authors.lastname: Zhang
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D004266:DNA Virus Infections / Q000235:genetics / Q000276:immunology / Q000662:veterinary / Q000821:virology; D005393:Fish Diseases / Q000235:genetics* / Q000276:immunology / Q000382:microbiology / Q000821:virology; D029941:Fish Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D005412:Flatfishes; D015870:Gene Expression; D006023:Glycoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000502:physiology; D016905:Gram-Negative Bacterial Infections / Q000235:genetics / Q000276:immunology / Q000382:microbiology / Q000662:veterinary; D006094:Gram-Positive Bacteria / Q000502:physiology; D016908:Gram-Positive Bacterial Infections / Q000235:genetics / Q000276:immunology / Q000382:microbiology / Q000662:veterinary; D007497:Iridoviridae / Q000502:physiology; D008074:Lipoproteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D010802:Phylogeny; D016415:Sequence Alignment
chemical_list: D029941:Fish Proteins; D006023:Glycoproteins; D008074:Lipoproteins; C051928:lipoprotein-associated coagulation inhibitor; C021240:tissue-factor-pathway inhibitor 2
keywords: Antibacterial activity; C-terminal peptides; Cynoglossus semilaevis; Expression; TFPI-1; TFPI-2