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xiaowanzi9新虫 (正式写手)
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Finally, we investigated direct effects of fumaric acid esters on the inhibitor of Nrf2, Keap1 in vitro. Mass spectroscopy revealed a covalent modification at the cysteine residue 151 of the Keap1 protein after monomethylfumarate treatment (Fig. 5G). The matched Keap1 peptides (peptide score 47) from a SpectrumMill database search covered 95% of the rat Keap1 protein sequence. After stimulation with monomethylfumarate, a 130 Da mass increase of the Keap1 tryptic peptide CVLHVMNGAVMYQIDSVVR was observed. The mass increase of 130 Da is consistent with a Michael addition of a free cysteine sulphydryl group across the monomethylfumarate double bond. The MS/MS spectrum of the peptide confirms this modification at cysteine 151 (Fig. 5G). The assignment of 130 Da to the monomethylfumarate modification is consistent (within 0.14 mmu) with a chemical formula of C5H6O4 (130.02661 exp. and 130.02647 cal.). The ion intensity ratio of the modified and unmodified peptides are 69 and 31%, respectively. The monomethylfumarate induced 130 Da modification was also observed––to a much lower extent––in other cysteine-containing Keap1 peptides (Table 2). |
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genhunter
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xiaowanzi9: 金币+20 2015-05-25 16:26:12
xiaowanzi9: 金币+20 2015-05-25 16:26:12
| 最后，我们研究了体外反丁烯二酸脂对Nrf2抑止物Keap1的直接作用(或效应). 质谱显示Keap1位于151的半胱氨酸上的巯基在反丁烯二酸单甲脂处理后被共价修饰了(图 5G)。通过从Spectrum Mill数据库中覆盖95%大鼠Keap1蛋白顺序检索找到了几个多肽片段(多肽同源值为47)。通过用反丁烯二酸单甲脂模拟，Keap1胰酶降解多肽片段CVLHVMNGAVMYQIDSVVR分子量会增加130 Da. 分子量增加130 Da是由于一个游离巯基与反丁烯二酸单甲脂双键通过迈克尔加成后形成的。. MS/MS图谱确认修饰位于位于151的半胱氨酸上. 分子量增加130 Da推算出反丁烯二酸单甲脂基团与其化学式C5H6O4相吻合(精确到0.14mmu, 实测130.02661 ，理论值 130.02647)。经修饰和未经修饰的多肽离子峰值比例分别是69和31%。反丁烯二酸单甲脂也导致同样130 Da修饰在其他含半胱氨酸的Keap1多肽片段中也可观察到，但含量要低很多。(表二) |
2楼2015-05-25 15:50:31
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xiaowanzi9: 金币+30, 翻译EPI+1 2015-05-25 16:26:09
xiaowanzi9: 金币+30, 翻译EPI+1 2015-05-25 16:26:09
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Finally, we investigated direct effects of fumaric acid esters on theinhibitor of Nrf2, Keap1 in vitro. 最后，我们研究了 反丁烯二酸酸酯对Keap1体外，Nrf2的抑制剂的直接作用 Mass spectroscopy revealed a covalent modification at the cysteineresidue 151 of the Keap1 protein after monomethylfumarate treatment (Fig. 5G). The matched Keap1 peptides (peptide score 47) from a Spectrum Mill database search covered 95% of the rat Keap1 protein sequence. 质谱分析表明， 运用富马酸单甲酯进行处理后，会形成一种，对Keap1蛋白质的半胱氨酸残基151的共价修饰（如图5G）。通过光谱数据库搜索获得的与之匹配的Keap1多肽（肽分为47）， 覆盖了95%的老鼠Keap1蛋白质序列。 After stimulation with monomethylfumarate,a 130 Da mass increase of the Keap1 tryptic peptide CVLHVMNGAVMYQIDSVVR was observed. The mass increase of 130 Da is consistent with a Michael addition of a free cysteine sulphydryl group across the monomethylfumarate double bond. 运用富马酸单甲酯进行刺激后，可以观察到，Keap1胰蛋白酶肽，会产生130 Da级的质量增加。该130Da的质量增加，与自由半胱氨酸巯基集团杂交富马酸单甲酯双键，的迈克尔加成反应一致。 The MS/MS spectrum of the peptide confirms this modification at cysteine 151 (Fig. 5G). The assignment of 130 Da to the monomethylfumarate modification is consistent (within 0.14 mmu) with a chemical formula of C5H6O4 (130.02661 exp. and 130.02647 cal.). The ion intensity ratio of the modified and unmodified peptides are 69 and 31%, respectively. 缩氨酸的串联式质谱法，确定了半胱氨酸151（如图5G） 修饰。130 Da分配到富马酸单甲酯修饰，与（在0.14 mmu内的）C5H6O4的化学式 （130.02661 exp. and 130.02647 cal.）一致。修饰/未修饰的肽的相对离子强度比分别为69% 和 31%。 The monomethylfumarate induced 130 Da modification was also observed––to a much lower extent––in other cysteine-containing Keap1 peptides (Table 2). 同时，也可以观察到富马酸单甲酯有法的130 Da的修饰。在含其它半胱氨酸的Keap1多肽中，其修饰程度更低。（如表二所示） |
3楼2015-05-25 15:54:36
