Characterization of metalloproteinase-like activities in barnacle (Balanus amphitrite) nauplii.
The presence of extracellular matrix (ECM) degrading enzymes was investigated in naupliar stages of the barnacle Balanus amphitrite Darwin. The results of substrate gel-zymography and quantitative assays demonstrated that naupliar extracts contain several protease activities that are specific towards gelatin substrates; some caseinolytic activity was also detected. Substrate specificity was observed in all naupliar stages (II-VI). The gelatinolytic activities showed dependence on both Ca(2+) and Zn(2+) and inhibition by EDTA, EGTA, and 1,10-phenanthroline. Also Mg(2+) partially activated the enzymes, whereas Cd(2+), Cu(2+), Hg(2+) and Pb(2+) were inhibitory. The thermal denaturation profile was significantly different in the presence and absence of Ca(2+) and Zn(2+). Overall, the results indicate that the Ca(2+)/Zn(2+)-dependent gelatinase activities in barnacle nauplii belong to the subfamily of matrix metalloproteases. Barnacle larvae MMPs showed biochemical characteristics different from those of vertebrate MMPs but common to other gelatinases from marine invertebrates: they were unaffected by several protease inhibitors and insensitive to specific activators/inhibitors of vertebrate MMPs. The presence of MMP-like activities in different naupliar stages suggests a constitutive role for these enzymes in ECM remodeling during barnacle larvae growth and development.
Mannello
Ferdinando
F
Istituto di Istologia ed Analisi di Laboratorio, Facoltà di Scienze, Via Zeppi, Università di Urbino Carlo Bo, 61029 Urbino (PU), Italy. f.mannello@uniurb.it
Canesi
Laura
L
Faimali
Marco
M
Piazza
Veronica
V
Gallo
Gabriella
G
Geraci
Sebastiano
S
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002413:Cations, Divalent; D002614:Chelating Agents / Q000494:pharmacology; D004789:Enzyme Activation / Q000187:drug effects; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005109:Extracellular Matrix / Q000201:enzymology*; D005780:Gelatin / Q000378:metabolism; D018093:Gelatinases / Q000037:antagonists & inhibitors / Q000378:metabolism; D006863:Hydrogen-Ion Concentration; D007814:Larva / Q000201:enzymology; D061965:Matrix Metalloproteinase Inhibitors; D020782:Matrix Metalloproteinases / Q000378:metabolism*; D013379:Substrate Specificity; D013696:Temperature; D001468:Thoracica / Q000201:enzymology*; D015032:Zinc / Q000494:pharmacology
D002413:Cations, Divalent; D002614:Chelating Agents; D004791:Enzyme Inhibitors; D061965:Matrix Metalloproteinase Inhibitors; D005780:Gelatin; D018093:Gelatinases; D020782:Matrix Metalloproteinases; D015032:Zinc; D002118:Calcium