Demonstration in yeast of the function of BP-80, a putative plant vacuolar sorting receptor
Demonstration in yeast of the function of BP-80, a putative plant vacuolar sorting receptor
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DOI：
10.1105/tpc.13.4.781
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发表时间：
2001-04-01
期刊：
影响因子：
11.6
通讯作者：
Paris, N
中科院分区：
文献类型：
--
作者：
Humair, D;Felipe, DH;Paris, N
BP-80，后来更名为VSRPS-1，是一种假定的受体，参与将蛋白质如糊粉蛋白原分选到裂解液泡，其N-末端结构域识别液泡分选决定簇。尽管迄今为止描述的所有VSRPS-1特征和体外结合特性都有利于其受体功能，但这种功能仍有待证实。在这里，我们使用绿色荧光蛋白（GFP）作为一个报告在酵母突变株缺陷为自己的液泡受体，Vps 10 p。通过表达VSRPS-1与融合到矮牵牛原糊粉蛋白的液泡分选决定簇的GFP，我们能够有效地将报告基因重定向到酵母液泡。VSRPS-1对单独的GFP或当与来自几丁质酶的另一种类型的植物液泡分选决定簇融合时是无效的。因此，植物VSRPS-1在体内与前糊粉蛋白液泡分选决定簇特异性相互作用，并且这种相互作用导致报告蛋白通过酵母分泌途径转运到液泡。这一发现证明了VSRPS-1受体的功能，但也强调了Vps 10 p及其植物等同物之间配体谱的差异。
BP-80, later renamed VSRPS-1, is a putative receptor involved in sorting proteins such as proaleurain to the lytic vacuole, with its N-terminal domain recognizing the vacuolar sorting determinant. Although all VSRPS-1 characteristics and in vitro binding properties described so far favored its receptor function, this function remained to be demonstrated. Here, we used green fluorescent protein (GFP) as a reporter in a yeast mutant strain defective for its own vacuolar receptor, Vps10p. By expressing VSRPS-1 together with GFP fused to the vacuolar sorting determinant of petunia proaleurain, we were able to efficiently redirect the reporter to the yeast vacuole. VSRPS-1 is ineffective on GFP either alone or when fused with another type of plant vacuolar sorting determinant from a chitinase. The plant VSRPS-1 therefore interacts specifically with the proaleurain vacuolar sorting determinant in vivo, and this interaction leads to the transport of the reporter protein through the yeast secretory pathway to the vacuole. This finding demonstrates VSRPS-1 receptor function but also emphasizes the differences in the spectrum of ligands between Vps10p and its plant equivalent.