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J Agric Food Chem. 2015 Feb 11;63(5):1488-95. doi: 10.1021/jf5046014. Epub 2015 Jan 28.

Purification and characterization of a novel lectin from Chinese leek seeds.

Author information

1
College of Biological Science and Technology, Fuzhou University , 2 Xue Yuan Road, University Town, Fuzhou, Fujian 350108, China.

Abstract

A novel lectin, CLSL, was purified from Chinese leek seeds by ion exchange chromatography on SP Sephadex C-25 and gel filtration chromatography on Sephadex G50. The lectin had a molecular weight of 23.6 kDa and was composed of two identical subunits linked by disulfide bonds, a conclusion based on SDS-PAGE under reducing and nonreducing conditions. CLSL was a glycoprotein with a carbohydrate content of 3.6%. It exerted potent agglutinating activity against rat red blood cells at a concentration of 8.9 μg/mL. Hemagglutination of rat erythrocytes was inhibited by d-fructose, mannitol, and sorbose at the concentration of 20 mM. The hemagglutinating activity of CLSL was maintained at 100 °C for 60 min and under acidic pH conditions but was lost at neutral and alkaline pH conditions. The hemagglutinating activity was stimulated by Ca(2+), Fe(2+), and Cu(2+) but inactivated by Ba(2+) at a concentration of 10 mM. Ba(2+)-mediated inactivation of CLSL was caused by CLSL conformational change induced by barium ions, according to the results of circular dichroism and fluorescence spectroscopy. Deconvolution of the CLSL circular dichroism indicated that it was an α-helical lectin with α-helix and β-fold contents of 35.8% and 8.6%, respectively. CLSL could also selectively inhibit cell proliferation.

KEYWORDS:

Chinese leek seeds; circular dichroism; fluorescence spectroscopy; lectin; purification

PMID:
25569192
DOI:
10.1021/jf5046014
[Indexed for MEDLINE]

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