Document Type : Research Paper
Inter University Centre for Bioscience and Department of Biotechnology and Microbiology, Kannur University, Thalassery Campus, Kannur 670661, India.
Background: The ethno-medical significance of Clerodendrum genus raises the interest towards the characterization of its seed lectin by inexpensive and most effective technique.
Objective: The focus of this study is the purification, characterization, and evaluation of the antioxidant and antiproliferative potential of a galactose-specific lectin from Clerodendrum infortunatum L. seeds.
Materials and Methods: The crude extract, homogenized in 6 volumes of the saline containing 10 mM β-mercaptoethanol was subjected to pigment removal by Toyopeal HW-55 column prior to ammonium sulfate fractionation (40-80 %).The crude protein extract was then loaded to the gel filtration column Sephadex G-200 followed by affinity chromatography using activated galactose coupled Sepharose-4B.
Results: The SDS-PAGE analysis showed a single band of about 30 kDa which further determined by MALDI-TOF analysis. The MALDI-TOF spectra revealed that Clerodendrum infortunatum lectin (CIL) is a homo-tetramer of 120 kDa consisting of four identical subunits of 30 kDa. The haemagglutination inhibition assay was done with purified lectin by many sugars, among which N-acetyl-D-galactosmine (NAG), D-galactose and lactose exhibited high inhibition. NAG showed the highest inhibition amongst the tested sugars, having the minimum inhibitory concentration of about 0.97 mM. The lectin exhibited a moderate antioxidant activity with an IC50 value of 6.1 ± 0.1 mg.mL-1 and induced cell death with IC50 of 82.8 μg.mL-1 against human gastric cancer cell line, AGS, indicated the potential of CIL for clinical and therapeutic applications.
Conclusion: The present study demonstrated the moderate ability of the CIL to inhibit the growth of human gastric cancer cells, AGS either by causing cytotoxic or anti-proliferative effects. Thus, CIL due to its remarkable properties may be considered as a potential bio-molecule in tumor research and glycobiology.