Speract. Purification and characterization of a peptide associated with eggs that activates spermatozoa.
A low molecular weight peptide (speract) associated with sea urchin eggs has been purified to apparent homogeneity by charcoal adsorption, DEAE-Sephacel chromatography, Bio-Gel P-2 filtration, and Dowex AG 50W-X4 chromatography. Gametes from 5000 female sea urchins were required for the isolation of approximately 9 mg of the peptide. The isolated peptide is homogenous based on [3H]acetic anhydride labeling, gel filtration, and reverse phase high pressure liquid chromatography. Speract is composed entirely of neutral and acidic amino acids with glycine as the major component, and it appears to have a blocked NH2 terminus based on its insensitivity to leucine aminopeptidase, its failure to react with dansyl chloride, and its chromatographic behavior on strong cation exchange resins. Speract is a potent stimulator of sea urchin sperm oxygen consumption, causing significant increases of sperm respiration rates at concentrations as low as 10(-12) M and producing 20-fold increases of oxygen consumption at maximal concentrations of 10(-8) M. Sperm cyclic GMP and cyclic AMP concentrations are also increased by speract, but concentrations of at least 10(-10) M and 10(-9) M are required for half-maximal elevations, respectively. The peptide, purified from Strongylocentrotus purpuratus eggs, also cross-reacts with spermatozoa from Lytechnis pictus sea urchins, suggesting that speract does not show species specificity. These results represent the first report of the purification of a peptide associated with eggs that may affect spermatozoa under natural conditions.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D001681:Biological Assay; D000242:Cyclic AMP / Q000378:metabolism; D006152:Cyclic GMP / Q000378:metabolism; D010450:Endopeptidases; D005260:Female; D008297:Male; D009842:Oligopeptides / Q000302:isolation & purification* / Q000502:physiology; D010063:Ovum / Q000502:physiology*; D010101:Oxygen Consumption / Q000187:drug effects; D012617:Sea Urchins; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*
D000596:Amino Acids; D009842:Oligopeptides; C028364:speract; D000242:Cyclic AMP; D010450:Endopeptidases; D006152:Cyclic GMP