Purification, molecular cloning, and characterization of glutathione S-transferases (GSTs) from pigmented Vitis vinifera L. cell suspension cultures as putative anthocyanin transport proteins.
Purification, molecular cloning, and characterization of glutathione S-transferases (GSTs) from pigmented Vitis vinifera L. cell suspension cultures as putative anthocyanin transport proteins.
复制标题
从色素葡萄葡萄葡萄葡萄酶（GSTS）的纯化，分子克隆和表征化的谷胱甘肽S-转移酶（GST）。
DOI：
10.1093/jxb/ern217
复制
发表时间：
2008
影响因子：
6.9
通讯作者：
Zhang W
中科院分区：
文献类型：
--
作者：
Conn S;Curtin C;Bézier A;Franco C;Zhang W
特定谷胱甘肽 S-转移酶 (GST) 的配体活性对于将花青素从细胞质转运到植物液泡是必需的。从 Vitis vinifera L. cv. 中纯化出 5 种 GST。通过谷胱甘肽亲和层析法进行 Gamay Fréaux 细胞悬浮培养。这些蛋白质经过了 Edman 测序和质谱指纹分析，所得片段与公共数据库中预测的 GST 进行了比对。克隆相应的编码序列，并在大肠杆菌中进行异源表达，用于确认 GST 活性。细胞悬浮液和葡萄浆果中这些候选 GST 基因和关键花青素生物合成途径基因（PAL、CHS、DFR 和 UFGT）对花青素积累的转录分析表明，它们分别与两个序列 VvGST1 和 VvGST4 呈强正相关。 VvGST1和VvGST4转运花青素的能力在异源玉米bronze-2互补模型中得到证实，为它们在葡萄细胞中作为花青素转运蛋白的功能提供了进一步的证据。此外，悬浮细胞和葡萄浆果中 VvGST1 和 VvGST4 的差异诱导表明这两种蛋白质之间存在功能差异。对这些候选配体的进一步研究可能会确定操纵植物和体外悬浮细胞中花青素积累的机制。
The ligandin activity of specific glutathione S-transferases (GSTs) is necessary for the transport of anthocyanins from the cytosol to the plant vacuole. Five GSTs were purified from Vitis vinifera L. cv. Gamay Fréaux cell suspension cultures by glutathione affinity chromatography. These proteins underwent Edman sequencing and mass spectrometry fingerprinting, with the resultant fragments aligned with predicted GSTs within public databases. The corresponding coding sequences were cloned, with heterologous expression in Escherichia coli used to confirm GST activity. Transcriptional profiling of these candidate GST genes and key anthocyanin biosynthetic pathway genes (PAL, CHS, DFR, and UFGT) in cell suspensions and grape berries against anthocyanin accumulation demonstrated strong positive correlation with two sequences, VvGST1 and VvGST4, respectively. The ability of VvGST1 and VvGST4 to transport anthocyanins was confirmed in the heterologous maize bronze-2 complementation model, providing further evidence for their function as anthocyanin transport proteins in grape cells. Furthermore, the differential induction of VvGST1 and VvGST4 in suspension cells and grape berries suggests functional differences between these two proteins. Further investigation of these candidate ligandins may identify a mechanism for manipulating anthocyanin accumulation in planta and in vitro suspension cells.
登录
查看更多内容
影响因子：
12.3
作者：
Dixon DP;Lapthorn A;Edwards R
通讯作者：
Edwards R
影响因子：
2.7
作者：
Conn, S;Zhang, W;Franco, C
通讯作者：
Franco, C
影响因子：
7.4
作者：
BILANG, J;STURM, A
通讯作者：
STURM, A
影响因子：
56.9
作者：
Alonso, JM;Stepanova, AN;Ecker, JR
通讯作者：
Ecker, JR
DOI：
10.1007/bf00265434
发表时间：
1992-06-01
期刊：
MOLECULAR & GENERAL GENETICS
影响因子：
--
作者：
BODEAU, JP;WALBOT, V
通讯作者：
WALBOT, V