Isolation, characterisation and cDNA sequencing of a new form of parvalbumin from carp semen.
Parvalbumins (Pv) are calcium-binding proteins present mainly in the muscle and nervous system where they act as a Ca(2+) buffer. Our previous work demonstrated the presence of Pv-I in carp semen and indicated the presence of a second Pv (Pv-II). The purpose of the present work was to identify, purify and determine the full-length cDNA sequence of Pv-II from carp testis. Pv-II from seminal plasma was purified by ion-exchange chromatography (IEC) and preparative electrophoresis, while the Pv-II from spermatozoa was purified by IEC, gel filtration and preparative electrophoresis. The purified Pv-II was submitted to an analysis of molecular mass, isoelectric point (pI), amino-acid sequence and oligomerisation ability. The amino-acid sequence was used to construct primers and obtain the full-length cDNA sequence of seminal-specific Pv-II from carp testis. Analysis of the cDNA sequence indicated that carp-testis Pv-II was distinct from carp-muscle parvalbumins. Pv-II was distinct from Pv-I regarding sequence, molecular mass and pI. Both parvalbumins had the ability to form oligomers or to bind to other proteins. Carp seminal plasma had a protective effect against parvalbumin oligomerisation. Pv-II underwent post-translational modification such as n-acetylation and cysteinylation. The present study is the first to report the full-length cDNA sequence of parvalbumin from carp testis.
Department of Gamete and Embryo Biology, Semen Biology Group, Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, 10-747 Olsztyn, Poland.
Mariola A
MA
Dietrich
Department of Gamete and Embryo Biology, Semen Biology Group, Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, 10-747 Olsztyn, Poland.
Błażej
B
Westfalewicz
Institute of Ichthyobiology and Aquaculture of Polish Academy of Sciences, Go?ysz, 43-520 Chybie, Poland.
Patrycja
P
Jurecka
Institute of Ichthyobiology and Aquaculture of Polish Academy of Sciences, Go?ysz, 43-520 Chybie, Poland.
Ilgiz
I
Irnazarow
Department of Gamete and Embryo Biology, Semen Biology Group, Institute of Animal Reproduction and Food Research of Polish Academy of Sciences, 10-747 Olsztyn, Poland.
Andrzej
A
Ciereszko
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002347:Carps / Q000235:genetics* / Q000378:metabolism; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000235:genetics*; D004591:Electrophoresis, Polyacrylamide Gel; D007525:Isoelectric Focusing; D007526:Isoelectric Point; D008297:Male; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010320:Parvalbumins / Q000737:chemistry / Q000235:genetics* / Q000302:isolation & purification*; D020033:Protein Isoforms; D011499:Protein Processing, Post-Translational; D012661:Semen / Q000737:chemistry*; D017422:Sequence Analysis, DNA; D020539:Sequence Analysis, Protein; D013094:Spermatozoa / Q000737:chemistry*
D018076:DNA, Complementary; D010320:Parvalbumins; D020033:Protein Isoforms