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Isolation, purification and characterization of an N-acetyl-D-lactosamine binding mitogenic and anti-proliferative lectin from tubers of a cobra lily Arisaema utile Schott

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Title Isolation, purification and characterization of an N-acetyl-D-lactosamine binding mitogenic and anti-proliferative lectin from tubers of a cobra lily Arisaema utile Schott
 
Creator Dhuna, Vikram
Dhuna, Kshitija
Singh, Jatinder
Saxena, Ajit Kumar
Agrawal, Satyam K.
Kamboj, Sukhdev Singh
 
Subject Chemical Sciences
 
Description Lectins are the carbohydrate-binding proteins of
non-immune origin which have been the subject of
intense investigation over the last few decades owing
to the variety of interesting biological properties.
Most of the lectins which have been purified and
characterized from plants have been obtained from
dicotyledons. In the present study a lectin was purified
from tubers of a monocot plant Arisaema utile
(AUL) Schott by affinity chromatography on asialofetuin-
linked amino activated silica beads. AUL
gave a single band in SDS-PAGE at pH 8.3 corresponding
to subunit Mr 13.5 kDa. The native molecular
mass of AUL was 54 kDa suggesting a homotetrameric
structure. AUL gave multiple bands in
isoelectric focusing and in native PAGE at pH 8.3.
AUL was inhibited by N-acetyl-D-lactosamine (Lac
NAc), a disaccharide and asialofetuin, a complex desialylated
serum glycoprotein. When treated with
denaturing agents, the lectin was stable in the presence
of urea (3 M), thiourea (4 M) and guanidine HCl
(4 M). AUL was a glycoprotein with a carbohydrate
content of 1.2%. Complete loss of activity was observed
upon modification of tryptophan residues of
the lectin. The activity was reduced to 25% after
modification of tyrosine. Chemical modification of
arginine, histidine, serine and cysteine residues of
AUL did not affect its activity. Using Far UV CD
spectra the estimated secondary structure was 37%
α-helix, 25% β-sheet and 38% random contributions.
The lectin showed potent mitogenic response towards
human lymphocytes. In vitro anti-proliferative assay
using 11 human cancer cell lines resulted in 50% inhibition
of six cell lines viz. SW-620, HCT-15,
SK-N-SH, IMR-32, Colo-205 and HT-29 at 38, 42, 43,
49, 50 and 89 μg/ml, respectively.
 
Date 2010
 
Type Article
PeerReviewed
 
Format application/pdf
 
Identifier http://iiim.csircentral.net/399/1/10.4236abb.2010.12012.pdf
Dhuna, Vikram and Dhuna, Kshitija and Singh, Jatinder and Saxena, Ajit Kumar and Agrawal, Satyam K. and Kamboj, Sukhdev Singh (2010) Isolation, purification and characterization of an N-acetyl-D-lactosamine binding mitogenic and anti-proliferative lectin from tubers of a cobra lily Arisaema utile Schott. Advances in Bioscience and Biotechnology, 01 (02). pp. 79-90. ISSN 2156-8456
 
Relation http://dx.doi.org/10.4236/abb.2010.12012
http://iiim.csircentral.net/399/