Identification of hepatic protein-protein interaction targets for betaine homocysteine S-methyltransferase.
Identification of hepatic protein-protein interaction targets for betaine homocysteine S-methyltransferase.
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DOI：
10.1371/journal.pone.0199472
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发表时间：
2018
期刊：
影响因子：
3.7
通讯作者：
Pajares MÁ
中科院分区：
文献类型：
--
作者：
Garrido F;Pacheco M;Vargas-Martínez R;Velasco-García R;Jorge I;Serrano H;Portillo F;Vázquez J;Pajares MÁ
蛋白质-蛋白质相互作用是调节酶功能的重要机制，允许代谢物通道，途径之间的串扰或翻译后修饰的引入。因此，许多高通量的研究已经进行了不同的病理生理环境下建立的蛋白质网络的光。令人惊讶的是，这种类型的信息是非常有限的酶的中间代谢，如甜菜碱同型半胱氨酸S-甲基转移酶，尽管其高肝abbraction和它的作用，同型半胱氨酸代谢。本文采用亲和纯化结合质谱和酵母双杂交两种方法，进一步揭示了甜菜碱同型半胱氨酸S-甲基转移酶在褐家鼠正常肝脏中的相互作用。共鉴定了131个非冗余的推定相互作用靶标，其中20个被选择用于通过免疫共沉淀进行进一步验证。通过两种不同方法验证的相互作用靶点包括：S-甲基甲硫氨酸高半胱氨酸甲基转移酶或甜菜碱高半胱氨酸甲基转移酶2、甲硫氨酸腺苷转移酶α1和α2、cAMP依赖性蛋白激酶催化亚基α、4-羟苯丙氨酸双加氧酶和醛缩酶B。网络分析确定了122个节点和165条边，以及有限数量的KEGG途径，包括：氨基酸的生物合成，半胱氨酸和甲硫氨酸代谢，剪接体和代谢途径。这些结果进一步扩展了肝脏蛋氨酸循环内的联系，并表明与其他代谢途径的假定交叉对话值得进一步研究。
Protein-protein interactions are an important mechanism for the regulation of enzyme function allowing metabolite channeling, crosstalk between pathways or the introduction of post-translational modifications. Therefore, a number of high-throughput studies have been carried out to shed light on the protein networks established under different pathophysiological settings. Surprisingly, this type of information is quite limited for enzymes of intermediary metabolism such as betaine homocysteine S-methyltransferase, despite its high hepatic abundancy and its role in homocysteine metabolism. Here, we have taken advantage of two approaches, affinity purification combined with mass spectrometry and yeast two-hybrid, to further uncover the array of interactions of betaine homocysteine S-methyltransferase in normal liver of Rattus norvegicus. A total of 131 non-redundant putative interaction targets were identified, out of which 20 were selected for further validation by coimmunoprecipitation. Interaction targets validated by two different methods include: S-methylmethionine homocysteine methyltransferase or betaine homocysteine methyltransferase 2, methionine adenosyltransferases α1 and α2, cAMP-dependent protein kinase catalytic subunit alpha, 4-hydroxyphenylpyruvic acid dioxygenase and aldolase b. Network analysis identified 122 nodes and 165 edges, as well as a limited number of KEGG pathways that comprise: the biosynthesis of amino acids, cysteine and methionine metabolism, the spliceosome and metabolic pathways. These results further expand the connections within the hepatic methionine cycle and suggest putative cross-talks with additional metabolic pathways that deserve additional research.
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影响因子：
64.5
作者：
Huttlin EL;Jedrychowski MP;Elias JE;Goswami T;Rad R;Beausoleil SA;Villén J;Haas W;Sowa ME;Gygi SP
通讯作者：
Gygi SP
影响因子：
5
作者：
BERGER, R;VANFAASSEN, H;SMITH, GPA
通讯作者：
SMITH, GPA
影响因子：
3.9
作者：
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通讯作者：
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作者：
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通讯作者：
Marcotte EM
用于深入分析人肝脏磷酸蛋白质组的酶辅助 RP-RPLC 方法
影响因子：
3.3
作者：
Bian, Yangyang;Song, Chunxia;Zou, Hanfa
通讯作者：
Zou, Hanfa