Heterologous expression of newly identified galectin-8 from sea urchin embryos produces recombinant protein with lactose binding specificity and anti-adhesive activity

Abstract

Galectin family members specifically bind beta-galactoside derivatives and are involved in different cellular events, including cell communication, signalling, apoptosis, and immune responses. Here, we report a tandem-repeat type galectin from the Paracentrotus lividus sea urchin embryo, referred to as Pl-GAL-8. The 933nt sequence encodes a protein of 34.73 kDa, containing the conserved HFNPRF and WGxExR motifs in the two highly similar carbohydrate-recognition domains (CRD). The three-dimensional protein structure model of the N-CRD confirms the high evolutionary conservation of carbohydrate binding sites. The temporal gene expression is regulated during development and transcripts localize at the tip of the archenteron at gastrula stage, in a subset of the secondary mesenchyme cells that differentiate into blastocoelar (immune) cells. Functional studies using a recombinant Pl-GAL-8 expressed in bacteria demonstrate its hemo-agglutinating activity on human red blood cells through the binding to lactose, as well as its ability in inhibiting the adhesion of human Hep-G2 cells to the substrate. The recent implications in autoimmune diseases and inflammatory disorders make Gal-8 an attractive candidate for therapeutic purposes. Our results offer a solid basis for addressing the use of the new Pl-GAL-8 in functional and applicative studies, respectively in the developmental and biomedical fields.

Sea Urchin Extracellular Proteins Design a Complex Protein Corona on Titanium Dioxide Nanoparticle Surface Influencing Immune Cell Behavior

“…Notably, the levels of these proteins in the sea urchin secretome exposed to TiO 2 NPs for 24 h was lower than that for the controls, perhaps because these proteins were sequestered on the particle surface. Both sea urchin toposome and nectin are glycoproteins (28, 35), while galectin-8 binds glycoproteins (36). Glycoproteins have good binding affinity to TiO 2 NP particle surface (37).…”

Subtle reproductive impairment through nitric oxide-mediated mechanisms in sea urchins from an area affected by harmful algal blooms

“…cDNA diluted 1:5 with H 2 O was used in Real Time qPCR experiments as described previously 24 . The data were normalized for the reference gene PlZ12-1 , which resulted to be the most suitable reference gene in previous studies 24 25 56 57 58 59 60 .…”

Carbonic anhydrases in development: morphological observations and gene expression profiling in sea urchin embryos exposed to acetazolamide

“…Pl -CAN was among the 10 occluded proteins identified, which were tightly bound to the mineral phase, and was present in both acetic acid-soluble and -insoluble matrices of the skeletal test tissues. By an in vitro crystallization assay, authors showed that this set of proteins has a shaping effect on the biocalcite formation, thus providing new information to the biologically controlled mineralization hypothesis [ 40 ]. The presence of Pl -CAN in adult tests was confirmed by WB using the same homologous polyclonal antibody used here to verify the presence of Pl -CAN in AZ-treated embryos ( figure 1 b this manuscript), indicating that Pl -CAN is a component of both embryonic and adult skeletal matrix and thus suggesting that it might have a similar behaviour in the adult and embryonic biomineralization processes.…”