Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm.
An essential initial step in fertilization in the sea urchin Strongylocentrotus purpuratus is an intracellular membrane fusion event in the sperm known as the acrosome reaction. This Ca2+-dependent, exocytotic process involves fusion of the membrane of the acrosomal vesicle and the plasma membrane. Recently, metalloendoproteases requiring divalent metals have been implicated in several Ca2+-dependent membrane fusion events in other biological systems. In view of the suggested involvement of Zn2+ in the sea urchin sperm acrosome reaction (Clapper, D.L., Davis, J.A., Lamothe, P.J., Patton, C., and Epel, D. (1985) J. Cell Biol. 100, 1817-1824) and the fact that Zn2+ is a metal cofactor for metalloendoproteases, we investigated the potential role of this protease in the acrosome reaction. A soluble metalloendoprotease was demonstrated and characterized in sperm homogenates using the fluorogenic protease substrate succinyl-alanine-alanine-phenylalanine-4-aminomethylcoumarin. The protease was inhibited by the metal chelators EDTA and 1,10-phenanthroline, and activity of the inactive apoenzyme could be reconstituted with Zn2+. The metalloendoprotease substrate and inhibitors blocked the acrosome reaction induced either by egg jelly coat or by ionophore, but had no effect on the influx of Ca2+. These observations suggest that inhibition occurs at a step independent of Ca2+ entry. Overall, the results of this study provide strong indirect evidence that the acrosome reaction requires the action of metalloendoprotease.
D000177:Acrosome / Q000201:enzymology*; D000818:Animals; D010450:Endopeptidases / Q000378:metabolism*; D005306:Fertilization; D005343:Fibrinolytic Agents / Q000378:metabolism; D007700:Kinetics; D008297:Male; D008666:Metalloendopeptidases; D008667:Metalloproteins / Q000378:metabolism; D012617:Sea Urchins; D013094:Spermatozoa / Q000201:enzymology* / Q000502:physiology*
D005343:Fibrinolytic Agents; D008667:Metalloproteins; D010450:Endopeptidases; D008666:Metalloendopeptidases