Dietrich Mariola A, Westfalewicz Błażej, Jurecka Patrycja, Irnazarow Ilgiz, Ciereszko Andrzej
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.
Institute of Ichthyobiology and Aquaculture of Polish Academy of Sciences, Go?ysz, 43-520 Chybie, Poland.
Reprod Fertil Dev. 2014 Oct;26(8):1117-28. doi: 10.1071/RD13181.
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.
小白蛋白（Pv）是主要存在于肌肉和神经系统中的钙结合蛋白，在这些组织中它们作为Ca(2+)缓冲剂发挥作用。我们之前的研究证明了鲤鱼精液中存在Pv-I，并表明存在第二种Pv（Pv-II）。本研究的目的是从鲤鱼睾丸中鉴定、纯化并确定Pv-II的全长cDNA序列。通过离子交换色谱（IEC）和制备电泳纯化精浆中的Pv-II，而通过IEC、凝胶过滤和制备电泳纯化精子中的Pv-II。对纯化后的Pv-II进行分子量、等电点（pI）、氨基酸序列和寡聚化能力分析。利用氨基酸序列设计引物，获得鲤鱼睾丸中精液特异性Pv-II的全长cDNA序列。对cDNA序列的分析表明，鲤鱼睾丸Pv-II与鲤鱼肌肉中的小白蛋白不同。Pv-II在序列、分子量和pI方面与Pv-I不同。两种小白蛋白都有形成寡聚体或与其他蛋白质结合的能力。鲤鱼精浆对小白蛋白寡聚化有保护作用。Pv-II经历了诸如N-乙酰化和半胱氨酸化等翻译后修饰。本研究首次报道了鲤鱼睾丸中小白蛋白的全长cDNA序列。
