arapidproteinsamplepreparationmethodbasedonorganicaqueousmicrowaveirradiationtechnique
文献类型：期刊论文
| 作者 | Wu Shuaibin2; Zhang Hongfei3; Yang Kaiguang2; Ma Junfeng1; Liang Zhen2; Zhang Lihua2; Zhang Yukui2 |
| 刊名 | sciencechinachemistry
 |
| 出版日期 | 2015-01-01 |
| 卷号 | 58期号:3页码:526 |
| ISSN号 | 1674-7291 |
| 英文摘要 | Fast and efficient sample preparation methods are a prerequisite for protein identification in bottom-up proteomics. Here, an innovative microwave irradiation sample preparation method was developed based on an optimized organic-aqueous solvent system for protein identification. Specifically, protein solutions containing high-concentration acetonitrile were subjected to 5 min microwave irradiation. After cooling down, trypsin was added and the digestion was performed with 30 s microwave irradiation, and the resulting peptides were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). A shortened processing time of only 5.5 min is needed with this method (more than 12 h is necessary in the traditional overnight protein sample preparation). Moreover, due to the absence of urea and other chaotropic reagents, the digests can be readily identified by MALDI-TOF MS. When an assessment of this method was performed by digesting a model protein BSA, 69% ± 3% sequence coverage corresponding to 47 ± 3 peptides was obtained, which shows better protein identification than that from the standard overnight protein sample preparation method (51% ± 2% sequence coverage and 23 ± 1 peptides). Another model protein α-casein was used for the analysis of protein phosphorylation with the newly developed method that yielded 4 phosphopeptides with 8 phosphorylation sites, whereas 3 phosphopeptides with 2 phosphorylation sites were obtained from the traditional overnight approach. Moreover, the organic-aqueous microwave irradiation method provides effective digestion for proteins down to fmol. |
| 语种 | 英语 |
| 源URL | [http://cas-ir.dicp.ac.cn/handle/321008/174012]  |
| 专题 | 大连化学物理研究所_中国科学院大连化学物理研究所 |
| 作者单位 | 1.约翰·霍普金斯大学 2.中国科学院大连化学物理研究所 3.China National Tobacco Quality Supervision and Test Center |
| 推荐引用方式 GB/T 7714 | Wu Shuaibin,Zhang Hongfei,Yang Kaiguang,et al. arapidproteinsamplepreparationmethodbasedonorganicaqueousmicrowaveirradiationtechnique[J]. sciencechinachemistry,2015,58(3):526. |
| APA | Wu Shuaibin.,Zhang Hongfei.,Yang Kaiguang.,Ma Junfeng.,Liang Zhen.,...&Zhang Yukui.(2015).arapidproteinsamplepreparationmethodbasedonorganicaqueousmicrowaveirradiationtechnique.sciencechinachemistry,58(3),526. |
| MLA | Wu Shuaibin,et al."arapidproteinsamplepreparationmethodbasedonorganicaqueousmicrowaveirradiationtechnique".sciencechinachemistry 58.3(2015):526. |
入库方式： OAI收割
来源：大连化学物理研究所
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