Reinvestigation of aminoacyl-tRNA synthetase core complex by affinity purification-mass spectrometry reveals TARSL2 as a potential member of the complex.
Reinvestigation of aminoacyl-tRNA synthetase core complex by affinity purification-mass spectrometry reveals TARSL2 as a potential member of the complex.
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DOI：
10.1371/journal.pone.0081734
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Lee C
中科院分区：
文献类型：
--
作者：
Kim K;Park SJ;Na S;Kim JS;Choi H;Kim YK;Paek E;Lee C
二十种不同的氨基酰基-tRNA合成酶(ARs)将每种氨基酸与其同源tRNA连接起来。个体ARS还与神经疾病、癌症和自身免疫性疾病中的各种非规范活动有关。其中，8个ARS(D、EP、I、K、L、M、Q和RARS)与3个ARS相互作用的多功能蛋白(AIMP)一起组装成多合成酶复合体(MSC)。然而，MSC的细胞功能和全局拓扑结构仍不清楚。为了了解MSC内部的复杂相互作用，我们分别以AIMP1、AIMP2和KARS为诱饵蛋白进行了亲和纯化-质谱分析(AP-MS)。质谱学数据被输入SAINT软件，以区分真实的相互作用和背景污染物。在HEK 293T细胞中，每个诱饵中共有40,134,101个蛋白的圣概率大于0.9。不断发现形成复杂的ARS，如DARS、EPRS、IARS、KARS、LARS、MARS、QARS和RARS与每个诱饵相互作用。在KARS沉淀物中发现了AIMP2-DX2和AIMP1亚型2等变异体。对质谱学数据的相对富集度分析表明，TARSL2(苏氨酰-tRNA合成酶Like-2)高度富含ARS-core复合体。TARSL2与其他ARS核心复合体成分的免疫共沉淀进一步证实了这种相互作用。我们建议将TARSL2作为ARS核心复合体的一个新成分。
Twenty different aminoacyl-tRNA synthetases (ARSs) link each amino acid to their cognate tRNAs. Individual ARSs are also associated with various non-canonical activities involved in neuronal diseases, cancer and autoimmune diseases. Among them, eight ARSs (D, EP, I, K, L, M, Q and RARS), together with three ARS-interacting multifunctional proteins (AIMPs), are currently known to assemble the multi-synthetase complex (MSC). However, the cellular function and global topology of MSC remain unclear. In order to understand the complex interaction within MSC, we conducted affinity purification-mass spectrometry (AP-MS) using each of AIMP1, AIMP2 and KARS as a bait protein. Mass spectrometric data were funneled into SAINT software to distinguish true interactions from background contaminants. A total of 40, 134, 101 proteins in each bait scored over 0.9 of SAINT probability in HEK 293T cells. Complex-forming ARSs, such as DARS, EPRS, IARS, Kars, LARS, MARS, QARS and RARS, were constantly found to interact with each bait. Variants such as, AIMP2-DX2 and AIMP1 isoform 2 were found with specific peptides in KARS precipitates. Relative enrichment analysis of the mass spectrometric data demonstrated that TARSL2 (threonyl-tRNA synthetase like-2) was highly enriched with the ARS-core complex. The interaction was further confirmed by coimmunoprecipitation of TARSL2 with other ARS core-complex components. We suggest TARSL2 as a new component of ARS core-complex.
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影响因子：
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作者：
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