Isolation, characterization and cDNA sequencing of acrosin from turkey spermatozoa

• Published in: Comparative Biochemistry and Physiology Part B: Biochemistry and Molecular Biology 2010-09, Vol.157 (1), p.127-136
• Main Authors: Słowińska, Mariola, Olczak, Mariusz, Liszewska, Ewa, Wątorek, Wiesław, Ciereszko, Andrzej
• Format: Article
• Language: English
• Subjects: Acrosin; Acrosin - chemistry; Acrosin - genetics; Acrosin - isolation & purification; Acrosin - metabolism; Amidohydrolases - metabolism; Amino Acid Sequence; Animals; Base Sequence; cDNA; Chemical Phenomena; Chromatography, Gel; Cloning, Molecular; DNA, Complementary - genetics; Enzyme Precursors - metabolism; Gelatinases - metabolism; Humans; Male; Molecular Sequence Data; Semen; Sequence Analysis, DNA; Sequence Homology, Amino Acid; Spermatozoa - enzymology; Trypsin Inhibitor, Kazal Pancreatic - metabolism; Turkey; Turkeys - genetics
• ISSN: 1096-4959; 1879-1107
• DOI: 10.1016/j.cbpb.2010.05.011

Acrosin (EC 3.4.21.10) is serine proteinase localized in the sperm acrosome and considered to play an essential role in fertilization. In contrast to mammalian, there are only limited data concerning avian acrosin, mostly focused on the characterization of mature enzyme. In the present study, acrosin was isolated from turkey spermatozoa using gel filtration in the presence of 4 M urea at acidic pH. N-terminal Edman sequencing allowed the identification of the first 26 N-terminal amino acids: VVGGTEALHG SWPWIVSIQNPRFAGT. This sequence was used to construct primers and obtain a cDNA sequence from the testis. The amino acid sequence deduced from the cDNA shows that turkey acrosin is initially synthesized as prepro-protein with 19-residue signal peptide. This signal sequence is followed by a 327-residue sequence corresponding to the acrosin zymogen. Turkey proacrosin does not contain a proline-rich segment at the C-terminal portion. Mature turkey acrosin is a two-chain molecule consisting of light and heavy chains and was found to be glycoprotein. The proacrosin/acrosin system exists in turkey spermatozoa and this system can be activated similarly to that of mammals. The high value of association constant strongly suggests that acrosin activity in turkeys can be controlled by a seminal plasma Kazal inhibitor under physiological conditions.