Isolation and characterization of transferrin from common carp (Cyprinus carpio L) seminal plasma.
Transferrin (Tf) in fish is recognized as a component of non-specific humoral defense mechanisms against bacteria. It is a major protein of common carp seminal plasma but its structure and localization in carp testis is unknown. In this study we developed a simple and efficient three-step purification procedure consisting of affinity chromatography (Con A-Sepharose), hydrophobic interaction chromatography (Phenyl Sepharose) and gel filtration (Superdex 200). The molecular mass of Tf has been determined to be 73.6 kDa and isoelectric point 5.1. The peculiar characteristics of carp transferrin were the lack of carbohydrate component and binding of iron ions by only one functional iron-binding site. Western blot analysis revealed a strong similarity of carp seminal plasma Tf to carp blood Tf and Tf from seminal plasma of other cyprinids but a lower similarity to salmonid and percid fishes. Tf was localized to the blood vessels of the carp testis which strongly suggest that most Tf of carp seminal plasma originates from blood. In conclusion, seminal plasma Tf has a unique structure and is similar or identical to blood Tf.
Dietrich
Mariola A
MA
Department of Gamete and Embryo Biology, Semen Biology Group, Institute of Animal Reproduction and Food Research, 10-747 Olsztyn, Poland. mario@pan.olsztyn.pl
Zmijewski
Dominik
D
Karol
Halina
H
Hejmej
Anna
A
Bilińska
Barbara
B
Jurecka
Patrycja
P
Irnazarow
Ilgiz
I
Słowińska
Mariola
M
Hliwa
Piotr
P
Ciereszko
Andrzej
A
D000818:Animals; D002347:Carps / Q000276:immunology*; D004591:Electrophoresis, Polyacrylamide Gel; D007150:Immunohistochemistry / Q000662:veterinary; D007501:Iron / Q000276:immunology; D007526:Isoelectric Point; D008297:Male; D008970:Molecular Weight; D012661:Semen / Q000276:immunology*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000662:veterinary; D013737:Testis / Q000276:immunology*; D014168:Transferrin / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*
D014168:Transferrin; D007501:Iron