title: Isolation and characterization of an ovoinhibitor, a multidomain Kazal-like inhibitor from Turkey (Meleagris gallopavo) seminal plasma.
abstract: Turkey seminal plasma contains three serine proteinase inhibitors. Two of them, with low molecular masses (6 kDa), were identified as single-domain Kazal-type inhibitors responsible for regulating acrosin activity. Our experimental objective was to isolate and characterize the inhibitor with the high molecular weight from turkey seminal plasma. The inhibitor was purified using hydrophobic interaction and affinity chromatography. Pure preparations of the inhibitor were used for identification by mass spectrometry, for determination of physicochemical properties (molecular weight, pI, and content and composition of the carbohydrate component), for kinetic studies, and for antibacterial tests. Gene expression and immunohistochemical detection of the inhibitor were analyzed in the testis, epididymis, and ductus deferens. The inhibitor with a high molecular weight from turkey seminal plasma was identified as an ovoinhibitor, which was found in avian semen for the first time. The turkey seminal plasma ovoinhibitor was a six-tandem homologous Kazal-type domain serine proteinase inhibitor that targeted multiple proteases, including subtilisin, trypsin, and elastase, but not acrosin. Our results suggested that hepatocyte growth factor activator was a potential target proteinase for the ovoinhibitor in turkey seminal plasma. The presence of the ovoinhibitor within the turkey reproductive tract suggested that its role was to maintain a microenvironment for sperm in the epididymis and ductus deferens. The turkey seminal plasma ovoinhibitor appeared to play a significant role in an antibacterial semen defense against Bacillus subtilis and Staphylococcus aureus.
authors.affiliation: Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland m.slowinska@pan.olsztyn.pl.
authors.forename: Mariola
authors.initials: M
authors.lastname: Słowińska
authors.affiliation: Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland.
authors.forename: Ewa
authors.initials: E
authors.lastname: Liszewska
authors.affiliation: Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Nynca
authors.affiliation: In Vitro and Cell Biotechnology Laboratory, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland.
authors.forename: Joanna
authors.initials: J
authors.lastname: Bukowska
authors.affiliation: Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Hejmej
authors.affiliation: Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bilińska
authors.affiliation: The Veterinary Diagnostic Laboratory SLW BioLab, Ostróda, Poland.
authors.forename: Jarosław
authors.initials: J
authors.lastname: Szubstarski
authors.affiliation: Department of Poultry Science, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Kozłowski
authors.affiliation: Department of Poultry Science, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
authors.forename: Jan
authors.initials: J
authors.lastname: Jankowski
authors.affiliation: Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Ciereszko
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015387:Egg Proteins, Dietary / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D015180:Electrophoresis, Gel, Two-Dimensional; D008297:Male; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017434:Protein Structure, Tertiary; D012661:Semen / Q000737:chemistry; D029589:Seminal Plasma Proteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D015842:Serine Proteinase Inhibitors / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification*; D014422:Turkeys / Q000378:metabolism
chemical_list: D015387:Egg Proteins, Dietary; D029589:Seminal Plasma Proteins; D015842:Serine Proteinase Inhibitors; C004224:ovoinhibitor
keywords: antibacterial activity; domestic animal reproduction; expression; male reproductive tract; ovoinhibitor; seminal plasma; turkey