標題: | Rapid enrichment of phosphopeptides and phosphoproteins from complex samples using magnetic particles coated with alumina as the concentrating probes for MALDI MS analysis |
作者: | Chen, Cheng-Tai Chen, Wei-Yu Tsai, Pei-Jane Chien, Kun-Yi Yu, Jau-Song Chen, Yu-Chie 應用化學系 應用化學系分子科學碩博班 Department of Applied Chemistry Institute of Molecular science |
關鍵字: | alumina;magnetic particles;phosphopeptides;phosphoproteins;MALDI MS |
公開日期: | 5-一月-2007 |
摘要: | In this study, we used nanocomposite magnetic particles coated with alumina as the affinity probes to selectively concentrate phosphorylated peptides and proteins from a low volume of sample solution. Tryptic digest products of phosphoproteins including alpha- and beta-caseins, human protein phosphatase inhibitor 1, nonfat milk, egg white, and a cell lysate were used as the samples to demonstrate the feasibility of this approach. In only 30 and 90 s, phosphopeptides and phosphoproteins sufficient for characterization by MALDI-MS were enriched by the particles, respectively. Proteins trapped on the particles could be directly digested on the particles. The same particles in the digest solution were employed for enrichment of phosphopeptides. We estimated the required time for performing the enrichment of phosphopeptides from complex samples and characterization by MALDI MS was within 5 min. A small volume (50 mu L) and a low concentration (5 x 10(-10) M) of tryptic digest product of a phosphoprotein sample could be dramatically enriched and characterized using this approach. |
URI: | http://dx.doi.org/10.1021/pr0604460 http://hdl.handle.net/11536/11238 |
ISSN: | 1535-3893 |
DOI: | 10.1021/pr0604460 |
期刊: | JOURNAL OF PROTEOME RESEARCH |
Volume: | 6 |
Issue: | 1 |
起始頁: | 316 |
結束頁: | 325 |
顯示於類別: | 期刊論文 |