Characterization of metalloproteinase-like activities in barnacle (Balanus amphitrite) nauplii

详细信息    查看全文

• 作者：Mannello ; Ferdinando ; Canesi ; Laura ; Faimali ; Marco ; Piazza ; Veronica ; Gallo ; Gabriella ; Geraci ; Sebastiano
• 关键词：Balanus amphitrite Darwin ; Barnacle ; Nauplii ; Extracellular matrix ; Zymography ; Gelatinase ; Matrix metalloproteinase
• 刊名：Comparative Biochemistry and Physiology, Part B
• 出版年：2003
• 出版时间：May, 2003
• 年：2003
• 卷：135
• 期：1
• 页码：17-24
• 全文大小：457 K

文摘

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 Ca²⁺ and Zn²⁺ and inhibition by EDTA, EGTA, and 1,10-phenanthroline. Also Mg²⁺ partially activated the enzymes, whereas Cd²⁺, Cu²⁺, Hg²⁺ and Pb²⁺ were inhibitory. The thermal denaturation profile was significantly different in the presence and absence of Ca²⁺ and Zn²⁺. Overall, the results indicate that the Ca²⁺/Zn²⁺-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.