Dioscorin, the major tuber storage protein of yam (Dioscorea batatas Decne) with carbonic anhydrase and trypsin inhibitor activities
Dioscorin, the major tuber storage protein of yam (Dioscorea batatas Decne) with carbonic anhydrase and trypsin inhibitor activities
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DOI：
10.1021/jf980738o
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发表时间：
1999-05-01
影响因子：
6.1
通讯作者：
Lin, YH
中科院分区：
文献类型：
--
作者：
Hou, WC;Liu, JS;Lin, YH
采用硫酸铵分馏、DE-52离子交换色谱和Sephadex G-75色谱柱对山药块茎贮藏蛋白薯蓣皂苷进行了分离纯化。SDS-PAGE后在非还原条件下发现两条蛋白带（82和28 kDa）；但在还原条件下只检测到一个条带（32 kDa）。28kda形式的n端前21个氨基酸为VEDEFSYIEGNPNGPENWGNL，与Conlan等（Plant Mol. Biol. 1995,28, 369-380）报道的薯蓣（Dioscorea cayeneasis Lam） cDNA中薯蓣皂苷的推断序列高度同源。Hewett-Emmett和tashan （Mol.系统发育学）。进化，1996,5,50 -77)提到，根据DNA比对，山药（D. cayenensis）的薯蓣皂苷与α -碳酸酐酶（α -ca）相关。在本报告中，我们发现纯化的diooscorin在使用碳酸氢钠作为底物时具有CA脱水活性，并且经过SDS-PAGE染色后具有CA活性。制备了一种针对甘薯贮藏蛋白胰蛋白酶抑制剂（TI）的多克隆抗体[j]。Lam var. Tainong 57)与薯蓣皂苷交叉反应，SDS-PAGE活性染色和胰蛋白酶抑制测定均显示TI活性。
Dioscorin, the tuber storage protein of yam (Dioscorea batatas Decne), was purified successively by ammonium sulfate fractionation, DE-52 ion exchange chromatography, and Sephadex G-75 column. Two protein bands (82 and 28 kDa) were found under nonreducing conditions after SDS-PAGE; but only one band (32 kDa) was detected under reducing conditions. The first 21 amino acids in the N-terminal region of the 28 kDa form were VEDEFSYIEGNPNGPENWGNL, which was highly homologous to deductive sequence of dioscorin from cDNA of another yam species (Dioscorea cayeneasis Lam) reported by Conlan et al. (Plant Mol. Biol. 1995, 28, 369-380). Hewett-Emmett and Tashian (Mol. Phylogenet. Evol. 1996, 5, 50 -77) mentioned that, according to DNA alignments, dioscorin from yam (D. cayenensis) was alpha-carbonic anhydrase (alpha-CA) related. In this report, we found that the purified dioscorin showed both CA dehydration activity using sodium, bicarbonate as a substrate and CA activity staining after SDS-PAGE. A polyclonal antibody, which was raised against trypsin inhibitor (TI), a storage protein of sweet potato (Ipomoea batatas [L.] Lam var. Tainong 57), cross-reacted with dioscorin, which also showed TI activity determined by both activity staining after SDS-PAGE and trypsin inhibition determination.