Hepatocyte growth factor activator is a potential target proteinase for Kazal-type inhibitor in turkey (Meleagris gallopavo) seminal plasma.
A peculiar characteristic of turkey seminal plasma is the increased activity of serine proteinases. It is of interest if the single-domain Kazal-type inhibitor controls the activity of turkey seminal plasma proteinases. Pure preparations of the Kazal-type inhibitor and anti-Kazal-type inhibitor monospecific immunoglobulin Gs were used as ligands in affinity chromatography for proteinase isolation from turkey seminal plasma. Gene expression and the immunohistochemical detection of the single-domain Kazal-type inhibitor in the reproductive tract of turkey toms are described. The hepatocyte growth factor activator (HGFA) was identified in the binding fraction in affinity chromatography. Hepatocyte growth factor activator activity was inhibited by the Kazal-type inhibitor in a dose-dependent manner. This protease was a primary physiological target for the single-domain Kazal-type inhibitor. Numerous proteoforms of HGFA were present in turkey seminal plasma, and phosphorylation was the primary posttranslational modification of HGFA. In addition to HGFA, acrosin was a target proteinase for the single-domain Kazal-type inhibitor. In seminal plasma, acrosin was present only in complexes with the Kazal-type inhibitor and was not present as a free enzyme. The single-domain Kazal-type inhibitor was specific for the reproductive tract. The germ cell-specific expression of Kazal-type inhibitors in the testis indicated an important function in spermatogenesis; secretion by the epithelial cells of the epididymis and the ductus deferens indicated that the Kazal-type inhibitor was an important factor involved in the changes in sperm membranes during maturation and in the maintenance of the microenvironment in which sperm maturation occurred and sperm was stored. The role of HGFA in these processes remains to be established.
Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland. Electronic address: m.slowinska@pan.olsztyn.pl.
Mariola
M
Słowińska
In Vitro and Cell Biotechnology Laboratory, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland.
Joanna
J
Bukowska
Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland.
Anna
A
Hejmej
Department of Endocrinology, Institute of Zoology, Jagiellonian University, Krakow, Poland.
Barbara
B
Bilińska
Department of Poultry Science, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Krzysztof
K
Kozłowski
Department of Poultry Science, Faculty of Animal Bioengineering, University of Warmia and Mazury in Olsztyn, Olsztyn, Poland.
Jan
J
Jankowski
Department of Gamete and Embryo Biology, Institute of Animal Reproduction and Food Research, Polish Academy of Sciences in Olsztyn, Olsztyn, Poland.
Andrzej
A
Ciereszko
D000176:Acrosin / Q000378:metabolism; D000818:Animals; D007150:Immunohistochemistry; D008297:Male; D010766:Phosphorylation; D011480:Protease Inhibitors / Q000378:metabolism*; D012661:Semen / Q000378:metabolism*; D012697:Serine Endopeptidases / Q000378:metabolism*; D014422:Turkeys / Q000378:metabolism*
D011480:Protease Inhibitors; C081092:HGF activator; D012697:Serine Endopeptidases; D000176:Acrosin
Acrosin; Expression; Hepatocyte growth factor activator; Seminal plasma; Single-domain Kazal-type inhibitor; Turkey