title: Characterization, expression and antibacterial properties of apolipoproteins A from carp (Cyprinus carpio L.) seminal plasma.
abstract: Apolipoproteins A are multifunctional proteins that, in addition to contributing to lipid metabolism and transport, are associated with the innate immune system in fish. Using a three step isolation procedure consisting of affinity chromatography on Blue-Sepharose, delipidation and reverse phase HPLC we isolated apolipoproteins from carp seminal plasma and identified them as ApoA-I and Apo-14 kDa. Moreover, we provided the full-length cDNA sequence of ApoA-I encoding 257 amino acids including a 18 amino acid signal peptide and a 4 amino acid propeptide. Apolipoproteins corresponded to the most abundant proteins in carp seminal plasma. Both ApoA-I and Apo-14 kDa were represented by several proteoforms that differ both in molecular mass and isoelectric point. The proteoforms of ApoA-I characteristic for seminal plasma were distinguished from those of blood. Carp seminal plasma ApoA-I and Apo-14 kDa showed a high immunologic similarity to their counterparts in carp blood and seminal plasma of other Cyprinid species. The mRNA expression analysis and immunohistochemical study suggest synthesis and secretion of ApoA-I and Apo-14 kDa in the fish reproductive tract and suggest a role in spermatogenesis and the stabilization of sperm membrane. Moreover, ApoA-I displayed bactericidal activity against Escherichia coli and bacteriostatic activity against Aeromonas hydrophila which suggests that ApoA-I is associated with innate immune system of the fish reproductive tract.
authors.affiliation: Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn, Department of Gamete and Embryo Biology, Poland. Electronic address: m.dietrich@pan.olsztyn.pl.
authors.forename: Mariola A
authors.initials: MA
authors.lastname: Dietrich
authors.affiliation: University of Veterinary Medicine in Hanover, Fish Disease Research Unit, Germany.
authors.forename: Mikołaj
authors.initials: M
authors.lastname: Adamek
authors.affiliation: Jagiellonian University in Krakow, Institute of Zoology, Poland.
authors.forename: Barbara
authors.initials: B
authors.lastname: Bilińska
authors.affiliation: Jagiellonian University in Krakow, Institute of Zoology, Poland.
authors.forename: Anna
authors.initials: A
authors.lastname: Hejmej
authors.affiliation: University of Veterinary Medicine in Hanover, Fish Disease Research Unit, Germany.
authors.forename: Dieter
authors.initials: D
authors.lastname: Steinhagen
authors.affiliation: Institute of Animal Reproduction and Food Research of Polish Academy of Sciences in Olsztyn, Department of Gamete and Embryo Biology, Poland.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Ciereszko
mesh_terms: D016980:Aeromonas hydrophila; D000704:Analysis of Variance; D000818:Animals; D001054:Apolipoproteins A / Q000032:analysis / Q000235:genetics* / Q000276:immunology*; D001424:Bacterial Infections / Q000276:immunology / Q000662:veterinary*; D015153:Blotting, Western; D002347:Carps; D002851:Chromatography, High Pressure Liquid; D003114:Colony-Forming Units Assay; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000235:genetics; D004926:Escherichia coli; D005393:Fish Diseases / Q000276:immunology* / Q000382:microbiology*; D007150:Immunohistochemistry; D008099:Liver / Q000378:metabolism; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012661:Semen / Q000737:chemistry*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D053719:Tandem Mass Spectrometry; D013737:Testis / Q000378:metabolism
chemical_list: D001054:Apolipoproteins A; D017931:DNA Primers; D018076:DNA, Complementary; D012333:RNA, Messenger
keywords: Antimicrobial properties; Apolipoprotein; Carp; Seminal plasma