Characterization and biological activities of lectin isolated from Pinellia ternata

Ruijuan Feng, Weibiao Zhang, Tao Xu

Institute of Bioengineering, Zhejiang Sci-Tech University, Hangzhou, China.

DOI: 10.4236/abc.2012.22014                

Abstract

In this paper, Pinellia ternata lectin (PTL) purified from 95% saturated ammonium sulfate precipitation by applying mannose-Sepharose 4B affinity column chromatography was first described. The mannose-Sepharose 4B affinity adsorption gel has already been designed and prepared before the experiments according the combinative characterization between lectin and mannose. Mass spectrometry analysis result shows that PTL is a glycoprotein with the molecular weight of 12.165 kD. It was conclude that PTL had strong agglutination effects on mouse red blood cells, and the minimum reaction concentration was 25 μg/ml according the Hemagglutination. Results of automatic amino acid analysis indicated that PTL mainly contained 15 varieties of amino acids, of which the minimum content was cysteine and aspartate was the maximum. Cell experiment results suggested that PTL of low concentrations (0.004 mg/ml, 0.02 mg/ml and 0.1 mg/ml) promoted HeLa cell proliferation, but the effect weakened with the concentrations and treated-time increasing. However, the HeLa cells pro- liferation was intensively inhibited by higher PTL concentrations (0.5 mg/ml and 1 mg/ml), and the effect increased in a dose and time dependent manner.

Keywords

Pinellia ternata Lectin; Mannose-Sepharose 4B Affinity Chromatography; Glycoprotein; Hela Cell; MTT Method

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Peumans, W.J. and Van Damme, E.J.M. (1995) Lectins as plant defense proteins. Plant Physiology, 109, 347-352. doi:10.1104/pp.109.2.347
[2]	Van Damme, E.J.M., Peumans, W.J., Barre, A. and Rougé, P. (1998) Plant lectins: A composite of several distinct families of structurally and evolutionary related proteins with diverse biological roles. Critical Reviews in Plant Sciences, 17, 575-692. doi:10.1080/07352689891304276
[3]	Guo, C.S., Yu, L.Y., Ge, W., Zhang, L. and Tang, C., et al. (2008) Research on extraction and hemagglutination effect of kidney bean phytohem agglutinin. Chinese Journal of Veterinary Drug, 42, 49-50.
[4]	Wang, K.Y. and Guo, M. (1993) Purification of lectin from Pinellia ternata. Chinese Biochemical Journal, 9, 544-547.
[5]	Zeng, Z.K., Wu, Q.Q. and Bao, J.K. (1995) Purification and characterization of cotton lectin. Acta Botanica Sinica, 37, 204-211.
[6]	Lin, Y.M. and Su, A.H. (2003) Isolation, purification and characterization of lectin from dictyophora indusiata fisch. Chinese Journal of Biochemistry and Molecular Biology, 19, 261-263.
[7]	Pan, K., Huang, B.Q. and Hou, X.W. (2007) Purification of a lectin from alocasia macrorrhiza and its toxic effect on four cultures insect cell line. Journal of Anhui Agricultural, 35, 5484-5485.
[8]	Li, J., Xu, X.C. and Bao, J.K. (2008) Effects of denaturation and chemical modification on lectin from phaseolus multiflorus. Willd. Natural Product Research and Development, 20, 787-792.
[9]	Gu, D.X. and Guo, Q.S. (1990) Study on biological characteristics in groups of Pinellia ternata (Thunb.) Breit. Journal of Nanjing Agricultural University, 13, 11-16.
[10]	Ge, X.Y. and Wu, H. (2009) Phytochemical properties and quality evaluation methods of Pinellia ternata. China Pharmaceuticals, 18, 3-5.
[11]	Li, B., Cheng, X.M., Zhou, Y.Y. and Lou, H.X. (2010) Progress on research of Pinellia ternata. Chinese Journal of Ethnomedicine and Ethnopharmacy, 19, 47-48.
[12]	Wang, Z.Q. and Li, B.C. (2009) Progress on the research of pharmacology of Pinellia ternata. Shanxi Medical Journal, 38, 65-67.
[13]	Xia, L.N. and Li, C.J. (1985) Study on the mechanism of anti-fertility effect and anti-early pregnancy of pinellin on mice. Journal of Fudan University (Medical Science), 12, 193-198.
[14]	Sun, G.X., Ding, S.S. and Qian, Y.J. (1992) Extraction, chemical analysis and effect on S-180 cell lines in mice of total protein of Pinellia pedatisecta schott. Journal of Fudan University (Medical Science), 1, 17-19.
[15]	Tao, Z.J., Xu, Q.Y., Wu, K.Z. and Lian S.H. (1981) Isolation and crystallization and biological activities and some chemical characteristics of pinellin. Acta Biochimica et Biophysica Sinica, 13, 77-82.
[16]	Wang, K.Y. and Guo M. (1993) Purification of lectin from Pinellia ternata. Chinese Biochemical Journal, 9, 544-547.
[17]	Wang Y.Q., Wu G.H., Lin X.F., He, L.P. and Huang, Z.L. (2009) Study on improving of polyacrylamide gel electrophoresis staining of polysaccharides and glycoproteins. Plant Physiology Communications, 45, 169-172.
[18]	Wang, H.W. (2008) Purification and N-terminal amino acid sequence analysis of one kind of protein with trypsin inhibitory activity in Pinellia ternata. Chinese Traditional Drug, 39, 1320-1323.
[19]	Zhu, M.W., Zhou, K.M. and Ding, S.S. (1999) Study on the effect of total protein of Pinellia pedatisecta schott on ovarian cancer cell lines and hematopoietic cells in human umbilical. Shanghai Medical University, 26, 455-456.
[20]	Huang, B.S. (2007) Study on apoptosis inducing effect of pinellin on human hepatoma carcinoma cells Bel-7402. Lishizhen Medicine and Materia Medica Research, 18, 1506.
[21]	Zhou, L., Wang, H.C., Zheng, Y.F. and Shi, M.R. (2010) Research on protein expression profile of human ovarian cancer cells SKOV3 treated by total protein of Pinellia pedatisecta schott by application of two-dimensional gel electrophoresis. Chinese Archives of Traditional Chinese Medicine, 28, 789-791.
[22]	Sun, C. (1981) Some biological properties of pinellin. Ac-ta Biochimica et Biophysica Sinica, 15, 1.
[23]	Tachibana, Y. and Kawanishi, K. (1992) Mitogenic activities in the protein fractions of crude drugs. Planta Medica, 58, 250-254.