Division of Biochemistry, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, SE-171 77 Stockholm, Sweden.
Department of Molecular Medicine and Haematology, University of the Witwatersrand Medical School, 2193 Johannesburg, South Africa.
J Biol Chem. 2013 Apr 5;288(14):10002-10011. doi: 10.1074/jbc.M112.445932. Epub 2013 Feb 14.
The human selenoprotein thioredoxin reductase 1 (TrxR1), encoded by the TXNRD1 gene, is a key player in redox regulation. Alternative splicing generates several TrxR1 variants, one of which is v3 that carries an atypical N-terminal glutaredoxin domain. When overexpressed, v3 associates with membranes and triggers formation of filopodia. Here we found that membrane targeting of v3 is mediated by myristoylation and palmitoylation of its N-terminal MGC motif, through which v3 specifically targets membrane rafts. This was suggested by its localization in cholera toxin subunit B-stained membrane areas and also shown using lipid fractionation experiments. Utilizing site-directed mutant variants, we also found that v3-mediated generation of filopodia is independent of the Cys residues in its redox active site, but dependent upon its membrane raft targeting. These results identify v3 as an intricately regulated protein that expands TXNRD1-derived protein functions to the membrane raft compartment.
人类硒蛋白硫氧还蛋白还原酶 1(TrxR1)由 TXNRD1 基因编码,是氧化还原调节的关键因子。选择性剪接产生了几种 TrxR1 变体,其中之一是携带非典型 N 端谷氧还蛋白结构域的 v3。当过度表达时,v3 与膜结合并触发纤毛的形成。在这里,我们发现 v3 的膜靶向是通过其 N 端 MGC 基序的豆蔻酰化和棕榈酰化介导的,通过这种方式 v3 特异性地靶向膜筏。这可以通过霍乱毒素亚基 B 染色的膜区域中的定位以及脂质分馏实验来证明。利用定点突变变体,我们还发现 v3 介导的纤毛生成不依赖于其氧化还原活性位点中的 Cys 残基,而是依赖于其膜筏靶向。这些结果表明 v3 是一种受精细调控的蛋白质,将 TXNRD1 衍生的蛋白质功能扩展到膜筏隔室。