Adaptation evolution and bioactivity of galectin from the deep sea Vesicomyidae clam Archivesica packardana

Hydrothermal vents and cold seep zones are two special habitats in the deep sea. These habitats are always dark, and have extreme temperatures (low or high), heavy metals and toxic substances (sulfide, methane). Vesicomyidae clams, which maintain endosymbionts in their gills, are common species in these two special zones and are thought to develop an efficacious immune system against unusual habitats. In the present study, a novel galectin (Apgalectin) was identified from the Vesicomyidae clam Archivesica packardana. The phylogenetic tree indicated that Apgalectin had two CRDs and was closely clustered with galectins from invertebrates, especially mollusks. A branch-site model showed that 9 positively selected sites (ω2 = 6.83950) were identified comparing to galectins from the Order Veneroida, implying a different function of Vesicomyidae galectins. A microbe binding assay showed that rApgalectin could bind to gram-positive bacteria, gram-negative bacteria and fungi. A PAMP binding assay indicated that Apgalectin could bind LPS, PGN, β-1,3-glucan, glucan from yeast and Poly I:C in dose-dependent manner. Apgalectin only agglutinated Micrococcus luteus and agglutination could be inhibited by galactose which demonstrated that Apgalectin might be involved in immune defense by recognizing and binding bacteria in a β-galactoside manner. Further experiments showed that Apgalectin might play an indirect effector role in the immune response because of its limited antibacterial spectrum. All analyses validated that Apgalectin from Archivesica packardana plays a variety of functions in immune responses and provided basal information for the immune study of deep-sea mollusks.