A rapid protein sample preparation method based on organic-aqueous microwave irradiation technique
文献类型：期刊论文
| 作者 | Wu, Shuaibin1,2; Zhang, Hongfei2; Yang, Kaiguang1; Ma, Junfeng3; Liang, Zhen1; Zhang, Lihua1; Zhang, Yukui1 |
| 刊名 | science china-chemistry
 |
| 出版日期 | 2015-03-01 |
| 卷号 | 58期号:3页码:526-531 |
| 关键词 | MALDI-TOF MS microwave organic-aqueous solvent protein identification digestion |
| 通讯作者 | 吴帅宾 |
| 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 a-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. |
| WOS标题词 | science & technology ; physical sciences |
| 学科主题 | 物理化学 |
| 类目[WOS] | chemistry, multidisciplinary |
| 研究领域[WOS] | chemistry |
| 关键词[WOS] | phase liquid-chromatography ; tandem mass-spectrometry ; maldi-tof-ms ; solvent systems ; efficient proteolysis ; enzymatic digestion ; silica microspheres ; formic-acid ; identification ; proteomics |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000350336500025 |
| 公开日期 | 2015-11-17 |
| 源URL | [http://159.226.238.44/handle/321008/143707]  |
| 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.Chinese Acad Sci, Dalian Inst Chem Phys, Natl Chromatog R&A Ctr, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China 2.China Natl Tobacco Qual Supervis & Test Ctr, Zhengzhou 450001, Peoples R China 3.Johns Hopkins Univ, Sch Med, Dept Biol Chem, Baltimore, MD 21205 USA |
| 推荐引用方式 GB/T 7714 | Wu, Shuaibin,Zhang, Hongfei,Yang, Kaiguang,et al. A rapid protein sample preparation method based on organic-aqueous microwave irradiation technique[J]. science china-chemistry,2015,58(3):526-531. |
| APA | Wu, Shuaibin.,Zhang, Hongfei.,Yang, Kaiguang.,Ma, Junfeng.,Liang, Zhen.,...&Zhang, Yukui.(2015).A rapid protein sample preparation method based on organic-aqueous microwave irradiation technique.science china-chemistry,58(3),526-531. |
| MLA | Wu, Shuaibin,et al."A rapid protein sample preparation method based on organic-aqueous microwave irradiation technique".science china-chemistry 58.3(2015):526-531. |
入库方式： OAI收割
来源：大连化学物理研究所
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