Nerve growth factor-mediated increases in protein methylation occur predominantly at type I arginine methylation sites and involve protein arginine methyltransferase 1
Nerve growth factor-mediated increases in protein methylation occur predominantly at type I arginine methylation sites and involve protein arginine methyltransferase 1
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DOI：
10.1002/jnr.10123
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发表时间：
2002-02-15
影响因子：
4.2
通讯作者：
Aletta, JM
中科院分区：
文献类型：
--
作者：
Cimato, TR;Tang, J;Aletta, JM
神经生长因子（NGF）特异性信号转导导致PC12细胞神经元分化过程中蛋白甲基化的变化（Cimato等a）。（1997）中华医学会医学杂志，32(3):391 - 391。在目前的工作中，我们证明，在ngf调节的蛋白质中，精氨酸甲基化比羧甲基化更普遍。I型蛋白精氨酸甲基转移酶（PRMT）活性导致底物蛋白中精氨酸末端胍二氮的不对称二甲基化，特别是蛋白质的甘氨酸和富含精氨酸（GAR）片段。几种GAR肽被用来测定PC12细胞提取物中甲基转移酶的活性，并与内源性细胞蛋白竞争PRMT活性。来源于纤维蛋白和核蛋白的肽，以及含有重复GRG基序的合成GAR肽，都能非常有效地阻断ngf调节的PC12细胞甲基化蛋白的体外甲基化。髓鞘碱性蛋白是II型PRMT的底物，选择性地抑制45 kDa蛋白，但在等摩尔浓度下，它是一种效果较差的总甲基化抑制剂。此外，利用纤维蛋白和核蛋白衍生肽检测ngf处理的PC12细胞匀浆中PRMT活性的升高。最后，从PC12细胞中免疫沉淀PRMT1首次证明了NGF激活的甲基转移酶，并表明PRMT1参与NGF信号转导。(C) 2002 Wiley-Liss, Inc。
Nerve growth factor (NGF)-specific signal transduction leads to changes in protein methylation during neuronal differentiation of PC12 cells (Cimato et a]. (1997] J. Cell Biol. 138:1089-1103). In the present work, we demonstrate that, among NGF-regulated proteins, arginine methylation is more prevalent than carboxylmethylation. Type I protein arginine methyltransferase (PRMT) activity produces asymmetric dimethylation of the terminal guanidinonitrogen of arginines in substrate proteins, particularly glycine and arginine-rich (GAR) segments of proteins. Several GAR peptides were used to assay for methyltransferase activity and to compete with endogenous cellular proteins for the PRMT activity in PC12 cell extracts. Peptides derived from fibrillarin and nucleolin, as well as a synthetic GAR peptide containing a repetitive GRG motif, are each extremely effective at blocking in vitro methylation of the NGF-regulated PC12 cell methylated proteins. Myelin basic protein, a substrate for type II PRMT, selectively inhibits a 45 kDa protein but is a much less effective inhibitor of total methylation at an equimolar concentration. In addition, the fibrillarin- and nucleolin-derived peptides were used to detect elevated PRMT activity in homogenates of NGF-treated PC12 cells. Finally, immunoprecipitation of PRMT1 from PC12 cells provides the first demonstration of an NGF-activated methyltransferase and implicates PRMT1 in NGF signal transduction. (C) 2002 Wiley-Liss, Inc.