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Purification, characterization and fine sugar specificity of a N-Acetylgalactosamine specific lectin from Adenia hondala

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Abstract

Plant lectins are gaining interest because of their interesting biological properties. Several Adenia species, that are being used in traditional medicine to treat many health ailments have shown presence of lectins or carbohydrate binding proteins. Here, we report the purification, characterization and biological significance of N-Acetyl galactosamine specific lectin from Adenia hondala (AHL) from Passifloraceae family. AHL was purified in a single step by affinity chromatography on asialofetuin Sepharose 4B column, characterized and its fine sugar specificity determined by glycan array analysis. AHL is human blood group non specific and also agglutinates rabbit erythrocytes. AHL is a glycoprotein with 12.5% of the carbohydrate, SDS-PAGE, MALDI-TOF-MS and ESI-MS analysis showed that AHL is a monomer of 31.6 kDa. AHL is devoid of DNase activity unlike other Ribosome inactivating proteins (RIPs). Glycan array analysis of AHL revealed its highest affinity for terminal lactosamine or polylactosamine of N- glycans, known to be over expressed in hepatocellular carcinoma and colon cancer. AHL showed strong binding to human hepatocellular carcinoma HepG2 cells with MFI of 59.1 expressing these glycans which was effectively blocked by 93.1% by asialofetuin. AHL showed dose and time dependent growth inhibitory effects on HepG2 cells with IC50 of 4.8 μg/ml. AHL can be explored for its clinical potential.

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Funding

SRI would like to thank Protein-Glycan Interaction Resource of the CFG (supporting grant R24 GM098791) and the National Center for Functional Glycomics (NCFG) at Beth Israel Deaconess Medical Center, Harvard Medical School (supporting grant P41 GM103694) for the glycan array analysis of AHL. The work was supported by the funding from University Grant Commission under UPE (F.NO.14 3/2012 (NS/PE) project.

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Authors and Affiliations

  1. Department of Studies in Biochemistry, Karnatak University, Dharwad, 580003, India

    Mamta Sharma, Prajna Hegde, Kavita Hiremath, Vishwanath Reddy H, Bale M. Swamy & Shashikala R. Inamdar

  2. Centre for Bioseparation Technology, VIT University, Vellore, 632014, India

    A. S. Kamalanathan

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  1. Mamta Sharma
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  2. Prajna Hegde
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  4. Vishwanath Reddy H
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  5. A. S. Kamalanathan
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  6. Bale M. Swamy
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  7. Shashikala R. Inamdar
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Contributions

Conceived and designed the experiments: SRI, AHL purification and characterization: MS, cell culture and Flow cytometry: PH and MS, glycan array and analyzed the data: SRI and VRH, DNase activity: KH, ESI/MS: KAS Contributed reagents/ materials/analysis tools: SRI, and BMS. Wrote the paper: SRI, BMS and MS.

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Correspondence to Shashikala R. Inamdar.

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This article does not contain any studies with human participants or animals performed by any of the authors.

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Sharma, M., Hegde, P., Hiremath, K. et al. Purification, characterization and fine sugar specificity of a N-Acetylgalactosamine specific lectin from Adenia hondala. Glycoconj J 35, 511–523 (2018). https://doi.org/10.1007/s10719-018-9843-6

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  • DOI: https://doi.org/10.1007/s10719-018-9843-6

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