Purification and partial peptide sequence analysis of the boar 32 kDa sperminogen

• Published in: Molecules and cells Vol. 12; no. 1; p. 107
• Main Authors: Yu, H, Yi, L S
• Format: Journal Article
• Language: English
• Published: United States 31-08-2001
• Subjects: Acrosin - chemistry; Acrosin - isolation & purification; Amino Acid Sequence; Animals; Chromatography, Gel; Electrophoresis, Polyacrylamide Gel; Enzyme Precursors - chemistry; Enzyme Precursors - isolation & purification; Male; Molecular Sequence Data; Peptide Fragments - chemistry; Seminal Plasma Proteins - chemistry; Seminal Plasma Proteins - isolation & purification; Spermatozoa - chemistry; Spermatozoa - enzymology; Swine
• ISSN: 1016-8478
• DOI: 10.1016/S1016-8478(23)17068-3

Boar 32 kDa sperminogen was purified from acid extracts of washed epididymal spermatozoa, and partial peptide sequence was determined. Boar sperminogen was purified from the acid extracts of boar spermatozoa by gel filtration through Sephadex G-75 column, followed by preparative SDS-PAGE. Gelatin zymographic analysis of the gel-filtered fractions showed that sperminogen was composed of three separate proteolytic bands. Among the three proteolytic bands, the 32 kDa sperminogen band which showed the strongest proteolytic activities upon activation was sliced out and eluted from the gel fragments. The eluted 32 kDa sperminogen was then subjected to peptide sequencing. Since the N-terminus of the 32 kDa sperminogen was blocked for peptide sequencing by Edman degradation method, the internal amino acid sequence of the sperminogen was obtained from the CNBr-digested peptides of sperminogen. The amino acid sequence of the analyzed peptide of the 32 kDa sperminogen showed 100% identity with that of proacrosin.