Varuna litterata (Fabricius) Lectin (VLL) Purification and Characterization

Author: Prakash Shoba S., Basil Rose, M.R. Venci Candida X., Anitha C. and Punitha A.

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Abstract

The carbohydrates are held together by the lectins which are glycoproteins. The apoptosis, involvement of pathogens, identification and tying of carbohydrates were some of the biological processes which happens. The adaptive and innate immune response, adaptive and innate immune response has an important part which is the binding features of universal ancient molecules called Sialic acid-binding lectins (SABLs). In this research, a novel sialic-binding lectin was isolated from the swimming crab Varuna litterata (Fabricius). The obstruction of growth and the abolishment of bacteria and fungi can be promoted by the accuracy of several carbohydrates present in the Lectins. The presence of reagents in biochemical research and the investigation of diagnosis were employed in the research of sialic acids for the treatment of pathogenic diseases and tumors which has attained a lot of interest in sialic acid-specific lectins. The molecular mass of the lectin was identified as 70 kDa on SDS-PAGE. In the current analysis, research was going on to isolate, identify and categorize the lectins from the haemolymph of agglutinin. To hinder agglutination, the potentiality of several glycoproteins and sugars (mono and oligosaccharides) was tested by making use of a hemagglutination inhibition (HAI) assay. After 1-hour, total inhibition of agglutination has been provided by the reciprocal of the lowest dilution of inhibitors which was shown by hemagglutination inhibition titer. The cross-adsorption assays have proceeded to identify whether there is the presence of single or multiple agglutinins in the hemolymph. Along with the desialylated rat erythrocytes, the agglutinin's preference for sialic acid is demonstrated by a decrease in hemagglutination activity. Moreover, a sepharose 4B column was connected with bovine submaxillary mucin which was triggered by CNBr to purify the lectin. Hemolymph agglutinin showed greater affinity towards the protease-treated rat erythrocytes and lesser affinity towards neuraminidase-treated rat erythrocytes than the native rat erythrocytes. As the O-acetyl sialic acid, a high preference was given for the inhibition of purified lectin by bovine submaxillary mucin and the non-inhibitory action of de-O-acetylated bovine submaxillary mucin. The results confirm the presence of an O-acetyl sialic acid-specific lectin in the hemolymph of the crab Varuna litterata respectively. The production of anti-tumor and antiviral drugs based on O-acetyl sialic acid-specific lectin in the hemolymph lectin of Varuna litterata may also have a significant utility in therapeutic industry.

Keywords

Varuna litterata, Sialic Acid, Lectin, Hemagglutination, Antigen, Agglutinin, Hemolymph, Glycoproteins, Rat Erythrocytes, Cross-Adsorption

Conclusion

The hemolymph of the freshwater crab Varuna literatta was purified lectin with a molecular weight of 70 kDa. The hemolymph lectin agglutinated with great avidity, the erythrocytes that exposed N-glycolyl neuraminic acid as the predominant epitope determinant. The isolated lectin was specific for the sugars D-galactose, N-acetyl–D-glucosamine, N-acetyl-Dmannosamine, trehalose and the sialoglycoproteins thyroglobulin, fetuin, and BSM. Sialic acid lectins or antibodies may be used as specialized sensors to examine the function of cell surface sugars during cellular development, differentiation, and malignant transformation. The purified hemolymph lectin against various mammalian erythrocytes was essentially the same as that of the crude serum; the ability of the lectin to agglutinate sialic acid-containing rat erythrocytes with great avidity suggests the possibility of the sialic acid specificity of the lectin. The hemolymph agglutinin was inhibited by N-acetyl sugars such as GluNAc, ManNAc, NeuAc and GalNAc, and sialic acid-containing glycoproteins, Bovine Submaxillary mucine> lactoferrin >holotransferrin = -acid glycoprotein > PSM > fetuin = apotransferrin. The outcome of this research strongly implies that V. literatta possesses a sialic acid-specific lectin in the hemolymph with a significant affinity for the O-acetyl group. This lectin could be employed as a useful diagnostic tool for determining O-acetyl NeuGc on the cell surfaces of microorganisms and tumour cells. The sialic acid specificity of the V. litterata lectin was confirmed by the inability of the lectin to agglutinate desialylated rat erythrocytes and the desialylated and de-O-acetylated BSM to inhibit hemagglutination. The specific binding property of salic acid and their respective functions since the identification of the binding site of the pathogens will be useful in medical and therapeutic research because of the complexity of glycoconjugates on the cell surface of pathogens.

References

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How to cite this article

Prakash Shoba S., Basil Rose, M.R. Venci Candida X., Anitha C. and Punitha A. (2023). Varuna litterata (Fabricius) Lectin (VLL) Purification and Characterization. Biological Forum – An International Journal, 15(1): 325-335.