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


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