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

Sci Rep. 2015 Dec 7:5:17665. doi: 10.1038/srep17665.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesives / metabolism*
  • Amino Acid Sequence
  • Animals
  • Cell Adhesion
  • Cloning, Molecular
  • Embryo, Nonmammalian / metabolism*
  • Embryonic Development / genetics
  • Galectins / chemistry
  • Galectins / metabolism*
  • Gene Expression Profiling
  • Gene Expression Regulation, Developmental
  • Hep G2 Cells
  • Humans
  • Lactose / metabolism*
  • Molecular Sequence Data
  • Paracentrotus / embryology*
  • Paracentrotus / genetics*
  • Phylogeny
  • Protein Binding
  • Protein Structure, Tertiary
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / isolation & purification
  • Recombinant Proteins / metabolism*
  • Sequence Analysis, Protein
  • Solubility
  • Structural Homology, Protein

Substances

  • Adhesives
  • Galectins
  • Recombinant Proteins
  • Lactose