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从半夏块茎和华山天南星块茎中分离并鉴定两种N-乙酰-D-乳糖胺特异性凝集素

Isolation and characterization of two N-acetyl-D-lactosamine specific lectins from tubers of Arisaema intermedium Blume and A. wallichianum Hook f.

作者信息

Kaur Manpreet, Singh Jatinder, Kamboj Sukhdev Singh, Singh Jagmohan, Kaur Amandeep, Sood S K, Saxena A K

机构信息

Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar 1430 05, India.

出版信息

Indian J Biochem Biophys. 2005 Feb;42(1):34-40.

Abstract

Two new lectins were purified from the tubers of Arisaema intermedium Blume and A. wallichianum Hook. f. (family: Araceae) by affinity chromatography on asialofetuin-linked amino activated silica beads. The bound lectins were eluted with 0.1 M glycine-HCl, pH 2.5. They gave a single band corresponding to subunit M(r) 13.4 kDa in SDS-PAGE, pH 8.3. On gel filtration chromatography, the lectins showed a M(r) of 51.2 kDa, suggesting a homotetrameric structure. Both the lectins gave a single peak on size exclusion HPLC and cation-exchange columns and a single band on PAGE, pH 4.5. However, like other monocot lectins, they gave multiple bands in isoelectric focusing and at PAGE 8.3. The lectins were inhibited by N-acetyl-D-lactosamine (LacNAc), a disaccharide and asialofetuin, a complex desialylated serum glycoprotein. They had no requirement for divalent metal ions i.e., Ca2+ and Mn2+ for their activity and were found to be mitogenic towards human lymphocytes. A. intermedium showed antiproliferative effect against various human cancer cell lines in vitro.

摘要

通过在去唾液酸胎球蛋白连接的氨基活化硅胶珠上进行亲和层析,从半夏(天南星科)和一把伞南星(天南星科)的块茎中纯化出两种新的凝集素。结合的凝集素用0.1 M甘氨酸 - HCl(pH 2.5)洗脱。在pH 8.3的SDS - PAGE中,它们呈现出一条对应于亚基M(r) 13.4 kDa的条带。在凝胶过滤层析中,凝集素的M(r)为51.2 kDa,表明其为同四聚体结构。两种凝集素在尺寸排阻HPLC和阳离子交换柱上均给出单峰,在pH 4.5的PAGE上给出单条带。然而,与其他单子叶植物凝集素一样,它们在等电聚焦和pH 8.3的PAGE中给出多条带。这些凝集素被二糖N - 乙酰 - D - 乳糖胺(LacNAc)和去唾液酸胎球蛋白(一种复杂的去唾液酸化血清糖蛋白)抑制。它们的活性不需要二价金属离子,即Ca2+和Mn2+,并且被发现对人淋巴细胞有促有丝分裂作用。半夏在体外对各种人类癌细胞系显示出抗增殖作用。

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