Interaction proteins of invertase and invertase inhibitor in cold-stored potato tubers suggested a protein complex underlying post-translational regulation of invertase
Interaction proteins of invertase and invertase inhibitor in cold-stored potato tubers suggested a protein complex underlying post-translational regulation of invertase
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冷藏马铃薯块茎中转化酶和转化酶抑制剂的相互作用蛋白表明转化酶翻译后调节背后存在蛋白质复合物
DOI：
10.1016/j.plaphy.2013.09.012
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发表时间：
2013-12-01
影响因子：
6.5
通讯作者：
Xie, Conghua
中科院分区：
文献类型：
--
作者：
Lin, Yuan;Liu, Jun;Xie, Conghua
液泡转化酶（VI）的活性对马铃薯冷甜化（CIS）起着至关重要的作用。VI活性的翻译后调节已被提出，其中涉及转化酶抑制剂（VIH），但VI和VIH之间的相互作用的机制尚未完全理解。为筛选VI和VIH的潜在配偶体，分别构建了抗CIS野生马铃薯Solanum berthautii和CIS敏感马铃薯品种AC 035 -01的cDNA文库，并进行酵母双杂交分析。将先前鉴定为与马铃薯CIS相关的StvacINV 1（马铃薯维斯之一）和StInvInh 2 B（马铃薯VIH之一）用作诱饵以筛选两个文库。通过阳性筛选和测序，筛选出StvacINV 1的27个潜在靶蛋白和StInvInh 2B的8个潜在靶蛋白。Kunitz型蛋白质抑制剂被StvacINV 1捕获在两个库中，它们之间的相互作用在烟草细胞中通过双分子荧光互补测定证实，加强了VI和VIH之间的基本相互作用。值得注意的是，蔗糖非发酵-1-相关的蛋白激酶1被捕获的诱饵，这表明蛋白质复合物可能是必要的转化酶活性的微调。本研究中阐明的靶蛋白为阐明VI活性可被微妙调节的机制提供了途径。(C)2013年Elsevier Masson SAS。All rights reserved.
The activity of vacuolar invertase (VI) is vital to potato cold-induced sweetening (CIS). A post-translational regulation of VI activity has been proposed which involves invertase inhibitor (VIH), but the mechanism for the interaction between VI and VIH has not been fully understood. To identify the potential partners of VI and VIH, two cDNA libraries were respectively constructed from CIS-resistant wild potato species Solanum berthaultii and CIS-sensitive potato cultivar AC035-01 for the yeast two-hybrid analysis. The StvacINV1 (one of the potato VIs) and StInvInh2B (one of the potato VIHs), previously identified to be associated with potato CIS, were used as baits to screen the two libraries. Through positive selection and sequencing, 27 potential target proteins of StvacINV1 and eight of StInvInh2B were clarified. The Kunitz-type protein inhibitors were captured by StvacINV1 in both libraries and the interaction between them was confirmed by bimolecular fluorescence complementation assay in tobacco cells, reinforcing a fundamental interaction between VI and VIH. Notably, a sucrose non-fermenting-1-related protein kinase 1 was captured by both the baits, suggesting that a protein complex could be necessary for fine turning of the invertase activity. The target proteins clarified in present research provide a route to elucidate the mechanism by which the VI activity can be subtly modulated. (C) 2013 Elsevier Masson SAS. All rights reserved.