Purification and characterization of a novel plant lectin from Pinellia ternata with antineoplastic activity
9 pages
English

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Purification and characterization of a novel plant lectin from Pinellia ternata with antineoplastic activity

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Description

A novel Pinellia ternata lectin was purified from the bulbs of a Chinese herb Pinellia ternata using a combination of hydrophobic chromatography and DEAE-ion exchange chromatography. The lectin was found to be a homodimer of 12093.3 Da subunits as determined by gel filtration and MS. Biochemical characterization of the lectin revealed the existence of a glycoprotein, which contains 3.22% neutral sugars. The N-terminal 10-amino acid sequence of the lectin, QGVNISGQVK, has not been reported for other lectins. The lectin had a special agglutinating activity with mouse erythrocytes at a minimum concentration of 8.0 ug/ml. The lectin was stable in the pH range of pH 5–12 and temperatures up to 80°C for 30 min. The results of MTT experiment showed that the lectin had significant effect towards tumor cells, the maximum inhibition of cell proliferation with Sarcoma 180, HeLa and K562 cell line were 85.2%, 74.6% and 59.4% respectively. Experimental therapy in vivo also showed that PTL apparently inhibited transplanted tumor in mice. Flow cytometric analysis demonstrated that PTL inhibited the proliferation of Sarcoma 180 in a time- and dose-dependent manner through inhibiting the transition of G 1 /S and subsequently inducing G 0 /G 1 cell cycle arrest. Thus, Pinellia ternata lectin displays a high potential for antitumor activity.

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Publié par
Publié le 01 janvier 2012
Nombre de lectures 46
Langue English

Extrait

Zuoet al. SpringerPlus2012,1:13 http://www.springerplus.com/content/1/1/13
a SpringerOpen Journal
R E S E A R C HOpen Access Purification and characterization of a novel plant lectin fromPinellia ternatawith antineoplastic activity 1 2*2 11 Zhenyu Zuo , Handong Fan, Xue Wang , Wei Zhouand Lingling Li
Abstract A novelPinellia ternatalectin was purified from the bulbs of a Chinese herbPinellia ternatausing a combination of hydrophobic chromatography and DEAEion exchange chromatography. The lectin was found to be a homodimer of 12093.3 Da subunits as determined by gel filtration and MS. Biochemical characterization of the lectin revealed the existence of a glycoprotein, which contains 3.22% neutral sugars. The Nterminal 10amino acid sequence of the lectin, QGVNISGQVK, has not been reported for other lectins. The lectin had a special agglutinating activity with mouse erythrocytes at a minimum concentration of 8.0 ug/ml. The lectin was stable in the pH range of pH 512 and temperatures up to 80°C for 30 min. The results of MTT experiment showed that the lectin had significant effect towards tumor cells, the maximum inhibition of cell proliferation with Sarcoma 180, HeLa and K562 cell line were 85.2%, 74.6% and 59.4% respectively. Experimental therapy in vivo also showed that PTL apparently inhibited transplanted tumor in mice. Flow cytometric analysis demonstrated that PTL inhibited the proliferation of Sarcoma 180 in a time and dosedependent manner through inhibiting the transition of G1/S and subsequently inducing G0/G1cell cycle arrest. Thus,Pinellia ternatalectin displays a high potential for antitumor activity. Keywords:Lectin,Pinellia ternata, Antineoplastic activity, Purification
Background Lectins are carbohydratebinding proteins of non immune origin with the ability to recognize special sugars existing on cell surfaces as a result of cell agglu tination (Peumans and Van Damme 1998). They are ubi quitous occurred in microorganisms, plants and animals and have attracted great interest due to their varied physiological roles in cell agglutination (Khan et al. 2007), antitumor (Liu et al. 2009, 2010), immunomodu latory (Rubinstein et al. 2004), antifungal (Herre et al. 2004) and antiviral effects (Wong and Ng 2005). Plant lectins have been extensively used for the detection, seg regation and description of glycoconjugates using their carbohydrate binding properties (Peumans and Van Damme 1998). In recent years, lectins are used to trigger vesicular transport into or across epithelial cells and
* Correspondence:fanhandong@whu.edu.cn 2 Institute of Microbiology, Jiangxi Academy of Sciences, Jiangxi 330029, China Full list of author information is available at the end of the article
used in food security management and diagnosis of dis ease. Furthermore, lectinmediated drugs have been acquired to target specific cells and some lectins with antiproliferative properties were isolated and character ized from different parts of the plant like seeds (Lin and Ng 2008), leaves (Park et al. 1997) and roots (Yan et al. 2010). Pinellia ternatais a traditional Chinese medicine which was used for the treatment of insomnia, eclampsia and termination of pregnancy for hundreds of years under the name of Banxia. It has been studied by many researchers due to the important discovery in the clinical application that the aqueous extract ofPinellia ternata contributed to the treatment of cervical carcinoma by smearing on the afflicted part (Lin et al. 2003). Much re search (Luo et al. 2000; Chen et al. 2003; Wu et al. 2011) has been done on indigenous compounds like polysac charide, astragaloside, isoflavonoids, triterpene, saponins and various trace elements inPinellasradix. However, not much information is available regarding the bio active proteins from this herb. Some researchers insisted
© 2012 Zuo et al.; licensee Springer. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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