Purification and characterization of a glycoprotein from Sipunculus nudus and its immune-enhancing activity to RAW 264.7 macrophages

Abstract

Sipunculus nudus, an edible marine invertebrate, has long been used as traditional Chinese medicine in folk remedies. In order to assess the immunoregulatory activity of glycoproteins in Sipunculus nudus and conduct a structure–activity relationship, a glycoprotein (SGP1) with molecular mass of 9.26 kDa was purified from Sipunculus nudus, and its chemical structure as well as immune-enhancing activity was investigated in this study. Structure analysis revealed that SGP1, a protein-dominate glycoprotein with O-glycosidic bonds, contained 92.8 % protein and 3.1 % saccharide. GC–MS result indicated that the saccharide moieties of SGP1 basically consisted of lyxose (Lyx), xylose (Xyl) as well as glucose (Glu) at a molar proportion of 0.87:4.16:1.36. The fourier transform infrared specoscopy (FT-IR) result proved that SGP1 have a typical characteristic of glycoprotein. Besides, circular dichroism (CD) result showed that SGP1 contained 4.1 % α-helix, 42.5 % β-sheet, 21.4 % β-turn, and 32.0 % random coil, indicating it’s mainly a β-sheet glycoprotein. The amino acid sequence of SGP1 shared a similarity to the Myohemerythrin (sp|Q5K473|HEMTM) with protein sequence coverage of 28.3 %. Moreover, the activity evaluation results showed that SGP1 exhibited significant immune-enhancing activity to the RAW 264.7 macrophages by promoting macrophages proliferation, enhancing phagocytic capacity, and simultaneously stimulating the secretions of nitric oxide (NO), interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) via NF-κB pathways. In this study, SGP1 as a novel glycoprotein had an obvious immune-enhancing activity to macrophages, and thus could be applied in the functional foods as a potential immunopotentiator for the hypoimmune population.