title: Purification and characterization of a Ca(2+)-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.
authors.affiliation: Department of Biochemistry and Molecular Biology, Faculty of Science, Rajshahi University, Rajshahi-6205, Bangladesh. rashelkabir@ru.ac.bd
authors.forename: Syed Rashel
authors.initials: SR
authors.lastname: Kabir
authors.forename: Md Abu
authors.initials: MA
authors.lastname: Zubair
authors.forename: Md
authors.initials: M
authors.lastname: Nurujjaman
authors.forename: Md Azizul
authors.initials: MA
authors.lastname: Haque
authors.forename: Imtiaj
authors.initials: I
authors.lastname: Hasan
authors.forename: Md Farhadul
authors.initials: MF
authors.lastname: Islam
authors.forename: Md Tanvir
authors.initials: MT
authors.lastname: Hossain
authors.forename: Md Anowar
authors.initials: MA
authors.lastname: Hossain
authors.forename: Md Abdur
authors.initials: MA
authors.lastname: Rakib
authors.forename: Mohammad Taufiq
authors.initials: MT
authors.lastname: Alam
authors.forename: Ranajit Kumar
authors.initials: RK
authors.lastname: Shaha
authors.forename: Md Tofazzal
authors.initials: MT
authors.lastname: Hossain
authors.forename: Yoshinobu
authors.initials: Y
authors.lastname: Kimura
authors.forename: Nurul
authors.initials: N
authors.lastname: Absar
mesh_terms: D000818:Animals; D002118:Calcium / Q000737:chemistry; D002286:Carcinoma, Ehrlich Tumor / Q000473:pathology*; D016923:Cell Death / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects*; D002645:Chickens; D002852:Chromatography, Ion Exchange; D003902:Detergents / Q000737:chemistry; D006023:Glycoproteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D006384:Hemagglutination / Q000187:drug effects; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000737:chemistry; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D051379:Mice; D030721:Nymphaea / Q000737:chemistry*; D035281:Plant Tubers / Q000737:chemistry*; D051381:Rats; D013050:Spectrometry, Fluorescence; D013696:Temperature; D014407:Tumor Cells, Cultured / Q000473:pathology
chemical_list: D003902:Detergents; D006023:Glycoproteins; D007477:Ions; D037102:Lectins; C569340:NNTL lectin, Nymphaea; D002118:Calcium