A membrane protein/signaling protein interaction network for Arabidopsis version AMPv2.
A membrane protein/signaling protein interaction network for Arabidopsis version AMPv2.
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拟南芥版本AMPV2的膜蛋白/信号蛋白相互作用网络。
DOI：
10.3389/fphys.2010.00024
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发表时间：
2010
影响因子：
4
通讯作者：
Frommer WB
中科院分区：
文献类型：
--
作者：
Lalonde S;Sero A;Pratelli R;Pilot G;Chen J;Sardi MI;Parsa SA;Kim DY;Acharya BR;Stein EV;Hu HC;Villiers F;Takeda K;Yang Y;Han YS;Schwacke R;Chiang W;Kato N;Loqué D;Assmann SM;Kwak JM;Schroeder JI;Rhee SY;Frommer WB
膜蛋白和可溶性部分之间的相互作用对于信号转导和调节营养转运至关重要。为了深入了解基于膜的相互作用组，从8,383个代表拟南芥膜和信号蛋白的目标列表中，将3,852个开放阅读框架（orf）克隆到gateway兼容载体中。利用基于交配的分裂泛素系统筛选490个拟南芥orf中潜在的蛋白相互作用（pppi）。在142种受体样激酶（RLKs）、72种转运蛋白、57种可溶性蛋白激酶和磷酸酶、40种糖基转移酶、95种不同功能的蛋白质和89种功能未知的蛋白质之间的二元机器人筛选中，检测出90,370种可能的ppi中的387种。二次筛选在179个蛋白之间确认343个（386个）pppi，产生无标度网络（r2 = 0.863）。142个跨膜RLKs中有80个检测呈阳性，鉴定出3个同质体，63个异质体，80个pppi与其他蛋白质。142个RLK相互作用体（包括RLK）中有31个先前被发现被磷酸化；因此相互作用物可以是各自rlk的底物。在主要的相互作用组数据库中，包括g蛋白偶联受体、磷脂酶C和AMT铵转运体的潜在相互作用物在内，本文所描述的pppi均未被报道。两个RLKs被认为是AMT1的相互作用物；1在拟南芥原生质体中使用分裂荧光素酶试验独立证实。这些RLKs可能参与氨依赖的c端磷酸化和铵吸收活性的调节。本文建立的机器人筛选方法将使真菌、植物和后生动物的膜蛋白相互作用的系统分析成为可能。
Interactions between membrane proteins and the soluble fraction are essential for signal transduction and for regulating nutrient transport. To gain insights into the membrane-based interactome, 3,852 open reading frames (ORFs) out of a target list of 8,383 representing membrane and signaling proteins from Arabidopsis thaliana were cloned into a Gateway-compatible vector. The mating-based split ubiquitin system was used to screen for potential protein–protein interactions (pPPIs) among 490 Arabidopsis ORFs. A binary robotic screen between 142 receptor-like kinases (RLKs), 72 transporters, 57 soluble protein kinases and phosphatases, 40 glycosyltransferases, 95 proteins of various functions, and 89 proteins with unknown function detected 387 out of 90,370 possible PPIs. A secondary screen confirmed 343 (of 386) pPPIs between 179 proteins, yielding a scale-free network (r2 = 0.863). Eighty of 142 transmembrane RLKs tested positive, identifying 3 homomers, 63 heteromers, and 80 pPPIs with other proteins. Thirty-one out of 142 RLK interactors (including RLKs) had previously been found to be phosphorylated; thus interactors may be substrates for respective RLKs. None of the pPPIs described here had been reported in the major interactome databases, including potential interactors of G-protein-coupled receptors, phospholipase C, and AMT ammonium transporters. Two RLKs found as putative interactors of AMT1;1 were independently confirmed using a split luciferase assay in Arabidopsis protoplasts. These RLKs may be involved in ammonium-dependent phosphorylation of the C-terminus and regulation of ammonium uptake activity. The robotic screening method established here will enable a systematic analysis of membrane protein interactions in fungi, plants and metazoa.
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影响因子：
7
作者：
Gisler, Serge M.;Kittanakom, Saranya;Stagljar, Igor
通讯作者：
Stagljar, Igor
影响因子：
14.9
作者：
Durek P;Schmidt R;Heazlewood JL;Jones A;MacLean D;Nagel A;Kersten B;Schulze WX
通讯作者：
Schulze WX
影响因子：
11.6
作者：
Chen, Zhongying;Noir, Sandra;Jones, Alan M.
通讯作者：
Jones, Alan M.
DOI：
10.1007/978-1-60761-765-5_24
发表时间：
2010-01-01
期刊：
PLANT DEVELOPMENTAL BIOLOGY: METHODS AND PROTOCOLS
影响因子：
--
作者：
Kato, Naohiro;Jones, Jason
通讯作者：
Jones, Jason
影响因子：
14.9
作者：
Käll L;Krogh A;Sonnhammer EL
通讯作者：
Sonnhammer EL