Proteomic profiling of tandem affinity purified 14-3-3 protein complexes in Arabidopsis thaliana

被引:161
作者
Chang, Ing-Feng [1 ,2 ,3 ]
Curran, Amy [1 ]
Woolsey, Rebekah [4 ]
Quilici, David [4 ]
Cushman, John C. [1 ]
Mittler, Ron [1 ]
Harmon, Alice [5 ,6 ]
Harper, Jeffrey F. [1 ]
机构
[1] Univ Nevada, Dept Biochem & Mol Biol, Reno, NV 89557 USA
[2] Natl Taiwan Univ, Inst Plant Biol, Taipei 10764, Taiwan
[3] Natl Taiwan Univ, Dept Life Sci, Taipei 10764, Taiwan
[4] Univ Nevada, Prote Core Facil, Reno, NV 89557 USA
[5] Univ Florida, Dept Bot, Gainesville, FL 32611 USA
[6] Univ Florida, Genet Inst, Gainesville, FL 32611 USA
基金
美国国家科学基金会;
关键词
14-3-3; Arabidopsis; Interactome; Kinase; Tandem affinity purification; MEMBRANE H+-ATPASE; PHOSPHATIDIC-ACID; PHOSPHOLIPASE-D; GLUTAMATE-RECEPTOR; 14-3-3-PROTEIN GF14-LAMBDA; K+ CHANNEL; PLANT; PHOSPHORYLATION; EXPRESSION; BINDING;
D O I
10.1002/pmic.200800445
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
In eukaryotes, 14-3-3 dimers regulate hundreds of functionally diverse proteins (clients), typically in phosphorylation-dependent interactions. To uncover new clients, 14-3-3 omega (At1g78300) from Arabidopsis was engineered with a "tandem affinity purification" tag and expressed in transgenic plants. Purified complexes were analyzed by tandem MS. Results indicate that 14-3-3 omega can dimerize with at least 10 of the 12 14-3-3 isoforms expressed in Arabidopsis. The identification here of 121 putative clients provides support for in vivo 14-3-3 interactions with a diverse array of proteins, including those involved in: (i) Ion transport, such as a K+ channel (GORK), a Cl- channel (CLCg), Ca2+ channels belonging to the glutamate receptor family (1.2, 2.1, 2.9, 3.4, 3.7); (ii) hormone signaling, such as ACC synthase (isoforms ACS-6, -7 and -8 involved in ethylene synthesis) and the brassinolide receptors BRI1 and BAK1; (iii) transcription, such as 7 WRKY family transcription factors; (iv) metabolism, such as phosphoenol pyruvate carboxylase; and (v) lipid signaling, such as phospholipase D (beta and gamma). More than 80% (101) of these putative clients represent previously unidentified 14-3-3 interactors. These results raise the number of putative 14-3-3 clients identified in plants to over 300.
引用
收藏
页码:2967 / 2985
页数:19
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