Purification and characterization of a Ca2+-dependent novel lectin from Nymphaea nouchali tuber with antiproliferative activities

Abstract

A lectin (termed NNTL) was purified from the extracts of Nymphaea nouchali tuber followed by anion-exchange chromatography on DEAE-cellulose, hydrophobic chromatography on HiTrap Phenyl HP and by repeated anion-exchange chromatography on HiTrap Q FF column. The molecular mass of the purified lectin was 27.0 ± 1.0 kDa, as estimated by SDS/PAGE both in the presence and in the absence of 2-mercaptoethanol. NNTL was an o-nitrophenyl β-D-galactopyranoside sugar-specific lectin that agglutinated rat, chicken and different groups of human blood cells and exhibited high agglutination activity over the pH range 5-9 and temperatures of 30-60 °C. The N-terminal sequence of NNTL did not show sequence similarity with any other lectin and the amino acid analysis revealed that NNTL was rich in leucine, methionine and glycine residues. NNTL was a glycoprotein containing 8% neutral sugar and showed toxicity against brine shrimp nauplii with an LC(50) value of 120 ± 29 μg/ml and exerted strong agglutination activity against four pathogenic bacteria (Bacillus subtilis, Sarcina lutea, Shigella shiga and Shigella sonnei). In addition, antiproliferative activity of this lectin against EAC (Ehrlich ascites carcinoma) cells showed 56% and 76% inhibition in vivo in mice at 1.5 and 3 mg·kg(-1)·day(-1) respectively. NNTL was a divalent ion-dependent glycoprotein, which lost its activity markedly in the presence of denaturants. Furthermore, measurement of fluorescence spectra in the presence and absence of urea and CaCl(2) indicated the requirement of Ca(2+) for the stability of NNTL.

Pea lectin inhibits growth of Ehrlich ascites carcinoma cells by inducing apoptosis and G2/M cell cycle arrest in vivo in mice

“…Multiplication of cancer cells was stopped by several kinds of plant lectin (Liu et al 2010) and due to the differences in their sugar specificity, each lectin exhibits differences in their antiproliferative effect against tumor cell lines. Several experiments were made to study the anticancer effect of lectins on different cancer cells (Liu et al 2010) but only a few reported against EAC cells (Ahmed et al 1988;Kabir et al 2011aKabir et al ,b, 2012Akev et al 2007). In the present study MTT assay was performed and the result showed that Pea lectin inhibited EAC cells proliferation in vitro in a dose-dependent manner.…”

“…Growth inhibition of EAC cells was also studied by using different lectins. Jackfruit lectin inhibited only 21.8, 40.2 and 57.5% of EAC cell growth at 50, 100 and 150 g/day respectively; Kaempferia rotunda lectin inhibited 51 and 67% of cell growth at 1.25 mg/kg/day and 2.5 mg/kg/day respectively; Trichosanthes cucumerina seed lectin (TCSL) showed 28 and 72% growth inhibition against EAC cells at 1 mg/kg/day and 2 mg/kg/day respectively and Nymphaea nouchali tuber lectin (NNTL) showed 56 and 76% growth inhibition against EAC cells at 1.5 mg/kg/day and 3.0 mg/kg/day respectively (Ahmed et al 1988;Kabir et al 2011aKabir et al ,b, 2012. In tumor-bearing mice, anemia occurs due to the decrease in RBC or hemoglobin percentage and increased the WBC.…”

“…They can specifically recognize and bind to various sugar structures, and thus trigger several important cellular processes (Sharon and Lis 1989;Sharon 2007). Several plant lectins have been shown to have antitumor activity and induce apoptosis in a series of tumor cell lines (Kabir et al 2011a(Kabir et al ,b, 2012Kim et al 2000;Liu et al 2010;De Mejía and Prisecaru 2005).…”

“…They are found in all kinds of organisms, including animals, plants, fungi, bacteria and viruses. Plant lectins represent a unique group of proteins with potent biological activities, such as agglutination, toxicity, anti-proliferation of cancer cells as well as having anti-fungal and anti-bacterial activities (Kabir et al 2011a(Kabir et al ,b, 2012Liu et al 2010;Sitohy et al 2007). They can specifically recognize and bind to various sugar structures, and thus trigger several important cellular processes (Sharon and Lis 1989;Sharon 2007).…”

A p‐Menth‐1‐ene‐4,7‐diol (EC‐1) from Eucalyptus camaldulensis Dhnh. Triggers Apoptosis and Cell Cycle Changes in Ehrlich Ascites Carcinoma Cells

“…Cell growth inhibition in vivo. To determine the cell growth inhibition properties of the compound, five groups of Swiss albino mice (n = 6) weighing 25 ± 4 g were used (Kabir et al, 2011). To assess the therapeutic evaluation, 1 × 10 6 EAC cells were inoculated into each group of mice on day 0.…”