title: A 53,000-Da esterase in Strongylocentrotus purpuratus semen is derived from phagocytic cells, not sperm.
abstract: Semen from the sea urchin Strongylocentrotus purpuratus contains sperm and a small volume (1%) of phagocytes, which often contain degraded sperm. A 53,000-Da esterase in the semen is inhibited by diisopropyl fluorophosphate, but not by soybean trypsin inhibitor (STI). Differential centrifugation experiments now reveal that 70% of the esterase activity (formerly described as a sperm protease precursor; Levine and Walsh, 1980) is associated with the phagocytes, which sediment more rapidly than the sperm. The 53K esterase is also present in spawned ovaries and testes. However, as previously reported, the sperm do contain an STI-inhibitable protease as shown by the digestion of [14C]lysozyme. Intact sperm exhibit STI-inhibitable hydrolytic activity toward N-alpha-[3H]benzoyl-L-arginine ethyl ester [3H]BAEE), but crude homogenates do not until they are extracted at pH 2.5 and fractionated by ion exchange chromatography. Although not obtained in pure form, the protease activity appears to migrate with a molecular weight of 20,000 (gel filtration). The protease and the esterase differ markedly in acid stability. After preincubation at pH 2.5 the protease still hydrolyzes [3H]BAEE, while the esterase is irreversibly inactivated. This last observation may explain an earlier interpretation (A. E. Levine and K. A. Walsh, 1980, J. Biol. Chem. 255, 4814-4820) that the 53K enzyme dissociated at pH 2.5 into two unequal subunits, one of which was the active protease. Since it has been shown that the contaminating phagocytes contribute most of the esterase activity of the semen, the occurrence of even a small number of nonsperm cells cannot be ignored in future investigations of sperm enzymes.
mesh_terms: D000818:Animals; D004950:Esterases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008970:Molecular Weight; D010447:Peptide Hydrolases / Q000378:metabolism; D010586:Phagocytes / Q000201:enzymology; D012617:Sea Urchins / Q000166:cytology / Q000201:enzymology*; D012661:Semen / Q000201:enzymology*; D013094:Spermatozoa / Q000201:enzymology*
chemical_list: D004950:Esterases; D010447:Peptide Hydrolases