Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm.

H. A. Farach, D. I. Mundy, W. J. Strittmatter, W. J. Lennarz

Research output: Contribution to journalArticle

35 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)5483-5487
Number of pages5
JournalJournal of Biological Chemistry
Volume262
Issue number12
StatePublished - Apr 25 1987

Fingerprint

Acrosome Reaction
Sea Urchins
Spermatozoa
Peptide Hydrolases
Fusion reactions
Metals
Membrane Fusion
Membranes
Alanine
Apoenzymes
Ionophores
Biological systems
Substrates
Cell membranes
Chelating Agents
Phenylalanine
Edetic Acid
Strongylocentrotus purpuratus
Intracellular Membranes
Fluorescent Dyes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Farach, H. A., Mundy, D. I., Strittmatter, W. J., & Lennarz, W. J. (1987). Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm. Journal of Biological Chemistry, 262(12), 5483-5487.

Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm. / Farach, H. A.; Mundy, D. I.; Strittmatter, W. J.; Lennarz, W. J.

In: Journal of Biological Chemistry, Vol. 262, No. 12, 25.04.1987, p. 5483-5487.

Research output: Contribution to journalArticle

Farach, HA, Mundy, DI, Strittmatter, WJ & Lennarz, WJ 1987, 'Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm.', Journal of Biological Chemistry, vol. 262, no. 12, pp. 5483-5487.
Farach, H. A. ; Mundy, D. I. ; Strittmatter, W. J. ; Lennarz, W. J. / Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm. In: Journal of Biological Chemistry. 1987 ; Vol. 262, No. 12. pp. 5483-5487.
@article{d3e2b07b124b445d8c35cddb2fe576fc,
title = "Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm.",
abstract = "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.",
author = "Farach, {H. A.} and Mundy, {D. I.} and Strittmatter, {W. J.} and Lennarz, {W. J.}",
year = "1987",
month = "4",
day = "25",
language = "English (US)",
volume = "262",
pages = "5483--5487",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "12",

}

TY - JOUR

T1 - Evidence for the involvement of metalloendoproteases in the acrosome reaction in sea urchin sperm.

AU - Farach, H. A.

AU - Mundy, D. I.

AU - Strittmatter, W. J.

AU - Lennarz, W. J.

PY - 1987/4/25

Y1 - 1987/4/25

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0023664261&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0023664261&partnerID=8YFLogxK

M3 - Article

VL - 262

SP - 5483

EP - 5487

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 12

ER -