Proteases from the Regenerating Gut of the Holothurian Eupentacta fraudatrix

Abstract

Four proteases with molecular masses of 132, 58, 53, and 47 kDa were detected in the digestive system of the holothurian Eupentacta fraudatrix. These proteases displayed the gelatinase activity and characteristics of zinc metalloproteinases. The 58 kDa protease had similar protease inhibitor sensitivity to that of mammalian matrix metalloproteinases. Zymographic assay revealed different lytic activities of all four proteases during intestine regeneration in the holothurian. The 132 kDa protease showed the highest activity at the first stage. During morphogenesis (stages 2–4 of regeneration), the highest activity was measured for the 53 and 58 kDa proteases. Inhibition of protease activity exerts a marked effect on regeneration, which was dependent on the time when 1,10-phenanthroline injections commenced. When metalloproteinases were inhibited at the second stage of regeneration, the restoration rates were decreased. However, such an effect proved to be reversible, and when inhibition ceased, the previous rate of regeneration was recovered. When protease activity is inhibited at the first stage, regeneration is completely abolished, and the animals die, suggesting that early activation of the proteases is crucial for triggering the regenerative process in holothurians. The role of the detected proteases in the regeneration processes of holothurians is discussed.

Manganese overload affects p38 MAPK phosphorylation and metalloproteinase activity during sea urchin embryonic development

“…auto-cleavage of a proenzyme which generates active forms with different molecular weights triggering a proteolytic cascade) increased in a time dependent-manner during development. This auto-cleavage activity was inhibited by EDTA, a versatile chelating agent of divalent ions with high affinity for calcium, known to prevent the endogenous auto-cleavage of calcium-dependent MMPs in echinoderms (Ranganathan et al, 2004;Lamash and Dolmatov, 2013). The seawater contains calcium ions at a high concentration (10 mM), thus we believe that, in analogy with what described in other sea urchin species (Quigley et al, 1993;Mayne and Robinson, 1998;Ranganathan et al, 2004), calcium ions play a key role in the regulation of most P. lividus MMPs.…”

Postembryonic Organogenesis of the Digestive Tube

“…Depending on the organization and composition, the extracellular matrix (ECM) can either promote regeneration or suppress it. The importance of connective tissue remodeling for initializing the regrowth of the viscera was demonstrated by experimental inhibition of matrix metalloproteases, which either slowed down or completely abolished gut regeneration in holothurians, depending on the treatment protocol (Lamash & Dolmatov, 2013; Quiñones, Rosa, Ruiz, & García-Arrarás, 2002). On the other side of the spectrum is the observation that if acute posttraumatic inflammation of the mammalian bowel leads to formation of the fibrotic scar, regeneration is inhibited and the normal intestinal tissue architecture is permanently altered, resulting in inability of the affected region to resume normal organ function (Speca et al, 2012).…”

Metabolite Profiling of Triterpene Glycosides of the Far Eastern Sea Cucumber Eupentacta fraudatrix and Their Distribution in Various Body Components Using LC-ESI QTOF-MS

“…E. fraudatrix is a well-studied species that is often used as a model organism in biological studies of various processes such as development [16,17], regeneration [18,19,20] and immunity [21,22,23]. The previous investigations of the sea cucumber E. fraudatrix have led to isolation of 37 triterpene glycosides comprising 15 non-sulfated tetraosides (cucumariosides A 1 –A 15 [24,25,26]), two non-sulfated triosides (cucumariosides B 1 and B 2 [27]), two non-sulfated pentaosides (cucumariosides C 1 and C 2 [28]), 4 sulfated tetraosides (cucumariosides G 1 –G 4 [29,30,31,32]), eight sulfated pentaosides (cucumariosides H, H 2 –H 8 [33,34,35]), two disulfated tetraosides (cucumariosides F 1 and F 2 [36]), and four disulfated pentaosides (cucumariosides I 1 –I 4 [37,38]).…”