DETECTION AND CHARACTERIZATION OF A VACUOLAR PROTEIN (VP24) IN ANTHOCYANIN-PRODUCING CELLS OF SWEET-POTATO IN SUSPENSION-CULTURE
DETECTION AND CHARACTERIZATION OF A VACUOLAR PROTEIN (VP24) IN ANTHOCYANIN-PRODUCING CELLS OF SWEET-POTATO IN SUSPENSION-CULTURE
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DOI：
10.1093/oxfordjournals.pcp.a078834
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发表时间：
1995-07-01
影响因子：
4.9
通讯作者：
YASUDA, H
中科院分区：
文献类型：
--
作者：
NOZUE, M;KUBO, H;YASUDA, H
从甘薯（Ipomoea batatas）花色苷产生细胞中分离纯化了含有大量蛋白质和花色苷的异戊醇。一个24 kDa的蛋白质（VP 24），确定为一个主要的蛋白质在分离的液泡，通过SDS-聚丙烯酰胺凝胶电泳检测。在暗培养的细胞中检测不到VP 24和花青素，但VP 24在开始光照后三天出现，并且VP 24的水平增加长达一周。高浓度的2，4-D显著抑制了VP 24和花青素的积累。这些结果表明，光诱导的VP 24的表达与花色素苷在液泡中的积累密切相关。VP 24在含有花青素苷的液泡的裂解物的超离心后作为不溶性的微红色沉淀物被回收。免疫印迹分析表明，这种液泡蛋白不同于甘薯块根的主要贮藏蛋白--孢子素。这些结果表明，VP 24是不是一个完整的蛋白质的液泡膜，但往往形成聚集体通过相互作用与花青素在提取过程中。VP 24可能参与形成液泡内色素结构，这些结构在培养的甘薯细胞中在含花青素的液泡中体内发育。
Vacuoles containing large amounts of protein and anthocyanin were isolated and purified from anthocyanin-producing cultured cells of sweet potato (Ipomoea batatas). A 24 kDa protein (VP24), identified as a major protein in the isolated vacuoles, was detected by SDS-polyacrylamide gel electrophoresis. Neither VP24 nor anthocyanin was detectable in dark-cultured cells, but VP24 appeared three days after the start of irradiation with light and the level of VP24 increased for up to a week. High concentrations of 2,4-D markedly inhibited the accumulation of both VP24 and anthocyanin. These results indicated that the expression of VP24, which was induced by exposure to light, was closely accompanied by the accumulation of anthocyanin in the vacuoles. VP24 was recovered as an insoluble reddish precipitate after ultracentrifugation of a lysate of anthocyanin-containing vacuoles. Immunoblot analysis indicated that this vacuolar protein was distinct from sporamin, a major storage protein of sweet potato tuberous root. These results suggest that VP24 is not an integral protein of the vacuolar membrane but tends to form aggregates via interactions with anthocyanin during extraction. VP24 may be involved in the formation of intravacuolar pigmented structures that develop in vivo in the anthocyanin-containing vacuoles of sweet potato cells in culture.