Isolation and biochemical characterization of the subunits of the rabbit sperm acrosome stabilizing factor.
The rabbit Acrosome Stabilizing Factor (ASF) is a glycoprotein synthesized in the corpus epididymis that demonstrates the ability to reversibly decapacitate sperm. Separation of the molecule into its individual subunits (92,000 Da and 38,000 Da) was accomplished via electroelution from polyacrylamide gels or via gel filtration on a Sephadex G-200 column in the presence of 0.1% sodium dodecyl sulfate. Column separation of the subunits revealed an entity of low molecular mass (500 daltons) associated with the ASF molecule. Amino acid compositional analysis of the subunits revealed the lack of cysteine and high glycine in the small subunit (38,000 Da) and high proline and glycine in the large subunit (92,000 Da). Lysine and aspartic acid were identified as the N-terminal amino acids for the large and small subunits, respectively. Identification of a 20 amino acid N-terminal sequence was accomplished for both of the subunits. Carbohydrate compositional analysis demonstrated that the small subunit contained N-asparagine-linked high mannose sugar chains while the large subunit contained N-asparagine-linked complex sugar chains. Endoglycosidase-H and N-Glycanase treatment of ASF indicated that the small subunit appears to contain four high mannose chains and the large subunit contains three complex chains.
D000177:Acrosome; D000595:Amino Acid Sequence; D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D002850:Chromatography, Gel; D006023:Glycoproteins / Q000302:isolation & purification*; D008297:Male; D008970:Molecular Weight; D010446:Peptide Fragments / Q000302:isolation & purification; D011487:Protein Conformation; D011817:Rabbits; D013075:Sperm Capacitation
C041608:ASF protein, Oryctolagus cuniculus; D000596:Amino Acids; D002241:Carbohydrates; D006023:Glycoproteins; D010446:Peptide Fragments