The specificity of sea urchin hatching enzyme (envelysin) places it in the mammalian matrix metalloproteinase family

Abstract

The sea urchin hatching enzyme (HEz) is a protease capable of dissolving the fertilization envelope that surrounds the embryo as a protective coat during early development. We have now purified a 37-kDa active enzyme from the supernatant fluid of hatched blastula medium of the species Hemicentrotus pulcherrimus. The purified enzyme was completely inhibited by alpha 2-macroglobulin and the chelating agents EDTA, EGTA, and 1,10-phenanthroline and was slightly inhibited by chymostatin and pepstatin, but was not inhibited by various other serine and thiol protease inhibitors. These results suggest that HEz is a metalloproteinase. Quantitative analyses of the products of HEz's action on various peptides revealed that the enzyme preferentially cleaved the peptide bonds on the amino side of bulky hydrophobic residues, -Leu, -Ile, and -Phe as well as -Tyr, in a similar but more limited fashion than thermolysin. Furthermore, although substance P was a good substrate of HEz, snake venom alpha-protease, and thermolysin, the analogue [Sar9]substance P was a poor substrate for HEz. This analogue was a good substrate for thermolysin and alpha-protease, but the scissile bonds were altered from those of substance P. The failure of HEz and alpha-protease to cleave the P1-P1 bond that meets the primary specificity is ascribed to the presence of an imino acid residue (proline or sarcosine) or the absence of any amino acid at the P2h or P3' position, in contrast to the simple P2' restriction of thermolysin.(ABSTRACT TRUNCATED AT 250 WORDS)

Purification and characterization of hatching enzyme from brine shrimp <italic>Artemia salina</italic>

“…Because of this complexity, activities with more specific biological relevance were sought. And it was found that the HE from shrimp [16] had not only proteolytic activity but also choriolytic activity, similar with those of echinoderms [1,5], fish [2,3,18], and amphibians [6,7,9,23]. In 1970, Yamagami [24] developed an effective method for HE activity assay based on the increase of 280 nm absorbance of chorionase digested turbid suspension of chorion fragments, and characterized the chorionase activity of HEs in Oryzias latipes successfully.…”

“…The biochemical properties of HE from many animal species, such as mammalian [4], avians [8], amphibians [6,7,9], teleostean [2,3,10,11], echinoderm [1,5], and insect [12], had been studied since 1980s. In invertebrates, HEs of sea urchin had been elucidated in terms of their functional properties, molecular structures, and gene structure [1,[13][14][15].…”

Structure, expression, and developmental function of early divergent forms of metalloproteinases in Hydra

“…A 30-res idue prodomain was identified based on its homo logy to the same region of Podocoryne metallopro teinase-1 (PMP-1) [21]. The existence of the pro domain was further supported by the N-terminal sequence of purified HMP-1, which suggested a proteolytic cleavage of secreted HMP-1 between Phe(51) and Lys (52). The processed HMP-1 wou ld have a predicted molecular mass of 27×10 3 , fit ting well with the size of purified HMP-1.…”

“…In contrast, relatively few MMPs have been identified in invertebrates. These invertebrate MMPs include envelysin from sea urchin [51][52][53], a Drosophila MMP [54]; and at least three separate MMPs from Caenorhabditis elegans [55]. Additional putative MMPs have been reported in plants such as soybean (Glycine max) [56], Arabidopsis thaliana [57], [58], and green alga [gamete lytic enzyme] [59].…”

On the Hatching Mechanism of Quail Embryos: Participation of Ectodermal Secretions in the Escape of Embryos from the Vitelline Membrane

“…Studies concerning the hatching mechanism have been conducted with sea urchins (Lepage and Gache, 1990;Nomura et al, 1991Nomura et al, , 1997Ghiglion et al, 1997), tunicate (D'Aniello et al, 1997), fish (Schoots et al, 1982;Yamagami et al, 1992;Yasumasu et al, 1994;Inohaya et al, 1995;Yamagami, 1996), amphibians (Yoshizaki, 1991;Yoshizaki and Yamasaki, 1991;Fan and Katagiri, 1997;Katagiri et al, 1997;Kitamura and Katagiri, 1998), and mammals (Perona and Wassarman 1986;Sawada et al, 1990). The common mechanism found in them is a disintegration of protecting envelopes by proteolytic hatching enzymes secreted by hatching gland cells (fish and amphibians) or temporarily participating anonymous cells (sea urchins and mammals).…”