Labeling primary amine groups in peptides and proteins with N-hydroxysuccinimidyl ester modified Fe3O4@SiO2 nanoparticles containing cleavable disulfide-bond linkers.
Labeling primary amine groups in peptides and proteins with N-hydroxysuccinimidyl ester modified Fe3O4@SiO2 nanoparticles containing cleavable disulfide-bond linkers.
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DOI：
10.1021/bc400165r
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发表时间：
2013-09-18
影响因子：
4.7
通讯作者：
Tarr, Matthew A.
中科院分区：
文献类型：
--
作者：
Patil, Ujwal S.;Qu, Haiou;Caruntu, Daniela;O'Connor, Charles J.;Sharma, Arjun;Cai, Yang;Tarr, Matthew A.
利用二硫键连接n -羟基琥珀酰亚胺（NHS）酯基团对超顺磁性二氧化硅包覆的氧化铁（Fe3O4@SiO2）纳米颗粒表面进行功能化，以建立一种标记肽/蛋白质中伯胺的方法。在没有任何额外的纯化步骤的情况下，使用磁铁分离NHS酯与伯胺基反应后形成的纳米颗粒标记的蛋白质/肽。然后用还原剂在二硫连接体上切割与肽/蛋白质结合的纳米颗粒片段。用LC-MS/MS分析标记的肽段，确定其序列和NHS酯标记位点。这种新方法可以表征天然牛血清白蛋白溶剂可及表面的赖氨酸残基。低成本、快速磁分离和对伯胺基团的特异性使NHS酯包被Fe3O4@SiO2纳米颗粒成为研究活细胞表面蛋白质的潜在标记探针。
The surface of superparamagnetic silica coated iron oxide (Fe3O4@SiO2) nanoparticles was functionalized with a disulfide bond linked N-hydroxysuccinimidyl (NHS) ester group in order to develop a method for labeling primary amines in peptides/proteins. The nanoparticle labeled proteins/peptides formed after NHS ester reaction with the primary amine groups were isolated using a magnet without any additional purification step. Nanoparticle moieties conjugated to peptides/proteins were then trimmed by cleavage at the disulfide linker with a reducing agent. The labeled peptides were analyzed by LC-MS/MS to determine their sequences and the sites of NHS ester labeling. This novel approach allowed characterization of lysine residues on the solvent accessible surface of native bovine serum albumin. Low cost, rapid magnetic separation, and specificity towards primary amine groups make NHS ester coated Fe3O4@SiO2 nanoparticles a potential labeling probe to study proteins on living cell surfaces.
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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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10.1016/j.saa.2010.04.004
发表时间：
2010-09-01
影响因子：
4.4
作者：
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通讯作者：
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