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马铃薯块茎和叶片中凝集素的纯化及性质;与马铃薯块茎酸性磷酸酶的相互作用

Purification and properties of lectin from potato tubers and leaves; interaction with acid phosphatase from potato tuber.

作者信息

Wierzba-Arabska E, Morawiecka B

机构信息

Institute of Biochemistry, Wroclaw University, Poland.

出版信息

Acta Biochim Pol. 1987;34(4):407-20.

PMID:3450103
Abstract

The haemagglutination activity of lectin, (STA) was determined during vegetation of potatoes (Solanum tuberosum var. Uran). Traces of haemagglutination activity were found in young stalks and first leaves. In the next leaves the maximal activity appeared after 6-8 weeks of vegetation and, simultaneously with the decrease in leaves it appeared in young tubers (at the time of their differentiation from stolons), reaching the maximum in the 18th week of vegetation. Lectins from potato tubers and leaves were extracted with acetate buffer, pH 3.6, salted out with ammonium sulphate, and purified alternatively by affinity chromatography on GlcNAc-Spheron or by adsorption on trypsinated and glutaraldehyde-fixed rabbit erythrocytes. Chromatography on Sephadex G-200 was used for preparative separation of isomeric forms. In the tubers four isolectins were found, differeing in molecular mass, pI values, content of sugars and haemagglutination activity. Two of these forms were isolated in homogeneous form: a heavy dimeric form, Mr 130,000-142,000 containing 59.6% of sugars (including 28.8% arabinose, 6.9% galactose, 16.5% glucose, 1.6% xylose and 3.1% mannose), and a light monomeric form. of Mr 21,000-22,000, containing 20% of neutral sugars (2.5% of pentoses and 17.5% of hexoses). From leaves, a single heavy dimeric lectin of Mr 136,000, containing 14.3% of neutral sugars was isolated. The effect of tuber isolectin on the activity, pH optima and heat-stability of the potato acid phosphatase has been described.

摘要

在马铃薯(Solanum tuberosum var. Uran)生长期间测定了凝集素(STA)的血凝活性。在幼茎和第一片叶子中发现了微量的血凝活性。在接下来的叶子中,最大活性在生长6 - 8周后出现,并且随着叶子中活性的降低,它出现在幼嫩块茎中(在它们从匍匐茎分化时),在生长第18周达到最大值。用pH 3.6的醋酸盐缓冲液从马铃薯块茎和叶子中提取凝集素,用硫酸铵盐析,然后交替通过在GlcNAc - Spheron上的亲和色谱或吸附在经胰蛋白酶处理和戊二醛固定的兔红细胞上进行纯化。使用Sephadex G - 200色谱进行异构体形式的制备性分离。在块茎中发现了四种异凝集素,它们在分子量、pI值、糖含量和血凝活性方面存在差异。其中两种形式以均一形式分离出来:一种重二聚体形式,Mr为130,000 - 142,000,含59.6%的糖(包括28.8%阿拉伯糖、6.9%半乳糖、16.5%葡萄糖、1.6%木糖和3.1%甘露糖),以及一种轻单体形式,Mr为21,000 - 22,000,含20%的中性糖(2.5%戊糖和17.5%己糖)。从叶子中分离出一种单一的重二聚体凝集素,Mr为136,000,含14.3%的中性糖。已经描述了块茎异凝集素对马铃薯酸性磷酸酶活性、最适pH值和热稳定性的影响。

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