Characterization of a metalloproteinase: a late stage specific gelatinase activity in the sea urchin embryo.
We have partially purified and characterized an 87 kDa gelatinase activity expressed in later stage sea urchin embryos. Cleavage activity was specific for gelatin and no cleavage of sea urchin peristome type I collagen, bovine serum albumin or casein was detected. Magnesium and Zn2+ inhibited the gelatinase and Ca2+ protected against inhibition. Ethylenediamine tetracetic acid, ethylenebisoxyethylenenitriol tetraacetic acid and 1,10-phenanthroline were inhibitory, suggesting that the gelatinase is a Ca(2+)- and Zn(2+)-dependent metalloproteinase. No inhibition was detected with serine or cysteine protease inhibitors and the vertebrate matrix metalloproteinase (MMP) inhibitor, Batimastat, was also ineffective. The vertebrate MMP activator p-aminophenylmercuric acetate was without effect. These results allow us to identify both similarities and differences between echinoderm and vertebrate gelatinases.
Robinson
J J
JJ
Department of Biochemistry, Memorial University of Newfoundland, St. John's, Canada. johnro@morgan.ucs.mun.ca
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002364:Caseins / Q000378:metabolism; D003094:Collagen / Q000737:chemistry / Q000378:metabolism; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004586:Electrophoresis / Q000379:methods; D004625:Embryo, Nonmammalian / Q000201:enzymology*; D005314:Embryonic and Fetal Development; D005775:Gastrula; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D008274:Magnesium / Q000494:pharmacology; D008666:Metalloendopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D051381:Rats; D012617:Sea Urchins / Q000196:embryology*; D012710:Serum Albumin, Bovine / Q000378:metabolism; D013379:Substrate Specificity; D013623:Tail / Q000737:chemistry; D015032:Zinc / Q000494:pharmacology
D002364:Caseins; D015853:Cysteine Proteinase Inhibitors; D012710:Serum Albumin, Bovine; D005780:Gelatin; D003094:Collagen; D018093:Gelatinases; D008666:Metalloendopeptidases; D008274:Magnesium; D015032:Zinc; D002118:Calcium