Accumulation of foreign polypeptides to rice seed protein body type I using prolamin portion sequences.
Accumulation of foreign polypeptides to rice seed protein body type I using prolamin portion sequences.
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DOI：
10.1007/s00299-016-2097-5
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发表时间：
2017-03
影响因子：
6.2
通讯作者：
Masumura T
中科院分区：
文献类型：
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作者：
Sasou A;Shigemitsu T;Morita S;Masumura T
水稻醇溶蛋白积累在内质网（ER）衍生的蛋白体，虽然保留在ER的保守序列没有得到证实。我们研究了必须在PB-Is中积累的醇溶蛋白的部分序列。 水稻种子醇溶蛋白积累在内质网（ER）衍生的蛋白体I型（PB-I），但在水稻醇溶蛋白多肽ER保留序列尚未得到证实。在这里，我们研究了在PB-I中积累所需的醇溶谷蛋白部分序列的长度。在水稻醇溶谷蛋白中，我们比较了13 a和13 b醇溶谷蛋白，因为这些醇溶谷蛋白的氨基酸序列非常相似，除了存在或不存在Cys残基。我们还产生和分析了转基因水稻表达几个醇溶蛋白部分序列-GFP融合蛋白。我们观察到，在13 α醇溶谷蛋白中，当部分序列从起始甲硫氨酸延伸超过第68个氨基酸残基时，醇溶谷蛋白部分序列-GFP融合蛋白在PB-1中积累。在13 b醇溶谷蛋白中，当部分序列从起始甲硫氨酸延伸超过第82个氨基酸残基时，醇溶谷蛋白部分序列-GFP融合蛋白在PB-1中积累。当在非还原或还原条件下提取这些融合蛋白时，PB-1中的13 α醇溶谷蛋白部分序列-GFP融合蛋白仅在还原条件下可溶。相反，13 b醇溶谷蛋白部分序列-GFP融合蛋白在非还原和还原条件下都是可溶的。这些结果表明，13 a醇溶谷蛋白在PB-Is中的积累与13 a醇溶谷蛋白多肽中二硫键和/或疏水性的形成有关，而13 b醇溶谷蛋白在PB-Is中的积累较少涉及二硫键的形成。本文的在线版本（doi：10.1007/s 00299 -016-2097-5）包含补充材料，可供授权用户使用。
Rice prolamins are accumulated in endoplasmic reticulum (ER)-derived proteins bodies, although conserved sequences retained in ER are not confirmed. We investigated portion sequences of prolamins that must accumulate in PB-Is. Rice seed prolamins are accumulated in endoplasmic reticulum (ER)-derived protein body type I (PB-I), but ER retention sequences in rice prolamin polypeptides have not been confirmed. Here we investigated the lengths of the prolamin portion sequences required for accumulation in PB-Is. Of the rice prolamins, we compared 13a and 13b prolamins because the amino acid sequences of these prolamins are quite similar except for the presence or absence of Cys-residues. We also generated and analyzed transgenic rice expressing several prolamin portion sequence-GFP fusion proteins. We observed that in 13a prolamin, when the portion sequences were extended more than the 68th amino acid residue from the initiating methionine, the prolamin portion sequence-GFP fusion proteins were accumulated in PB-Is. In 13b prolamin, when the portion sequences were extended by more than the 82nd amino acid residue from the initiating methionine, the prolamin portion sequence-GFP fusion proteins were accumulated in PB-Is. When those fusion proteins were extracted under non-reduced or reduced conditions, the 13a prolamin portion sequence-GFP fusion proteins in PB-Is were soluble under only the reduced condition. In contrast, 13b prolamin portion sequence-GFP fusion proteins were soluble under both non-reduced and reduced conditions. These results suggest that the accumulation of 13a prolamin in PB-Is is associated with the formation of disulfide bonds and/or hydrophobicity in 13a prolamin polypeptide, whereas the accumulation of 13b prolamin in PB-Is was less involved in the formation of disulfide bonds. The online version of this article (doi:10.1007/s00299-016-2097-5) contains supplementary material, which is available to authorized users.