Coupling hydrophilic interaction chromatography materials with immobilized Fe(3+) for phosphopeptide and glycopeptide enrichment and separation.
Coupling hydrophilic interaction chromatography materials with immobilized Fe(3+) for phosphopeptide and glycopeptide enrichment and separation.
复制标题
亲水相互作用层析材料与固定化Fe(3)偶联用于磷酸肽和糖肽的富集和分离
作者：
Simultaneous profiling of protein phosphorylation and glycosylation is very important to elucidate the bio-functions of these proteins. However, simultaneous enrichment of glyco- and phosphopeptides is the bottleneck in proteomics because of the low abundance of these species and ion suppression from non-modified peptides in mass spectrometry (MS). In this study, Fe3+ immobilized hydrophilic interaction chromatography (HILIC) materials (termed polySD-SiO2, recently reported in our lab) and polySD-SiO2 in the HILIC mode were employed for the simultaneous enrichment and subsequent separation of glyco- and phosphopeptides. The Fe3+ immobilized polySD-SiO2 could selectively enrich glycopeptides and phosphopeptides and the co-enriched peptides were further fractionated with polySD-SiO2 in the HILIC mode. With the established method, glyco- and phosphopeptides were well enriched and divided into two fractions even from tryptic digests of a-casein, fetuin and BSA at a molar ratio of 1 : 2 : 400. Application of the established method to HeLa cell lysate resulted in a total of 1903 phosphopeptides and 141 glycosylation sites. These results demonstrate that the established method could selectively and simultaneously enrich and fractionate glyco- and phosphopeptides from complex peptide mixtures. Simultaneous profiling of protein phosphorylation and glycosylation is very important to elucidate the bio-functions of these proteins.
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DOI：
10.1007/978-1-61779-319-6_23
发表时间：
2011-01-01
期刊：
NANOPROTEOMICS:METHODS AND PROTOCOLS
影响因子：
--
作者：
Hao, Piliang;Zhang, Huoming;Sze, Siu Kwan
通讯作者：
Sze, Siu Kwan
影响因子：
1.6
作者：
Kuroda, I;Shintani, Y;Furuno, M
通讯作者：
Furuno, M
影响因子：
4.4
作者：
Waniwan, Juanilita T.;Chen, Yi-Ju;Chen, Yu-Ju
通讯作者：
Chen, Yu-Ju
影响因子：
64.5
作者：
Chung JJ;Shim SH;Everley RA;Gygi SP;Zhuang X;Clapham DE
通讯作者：
Clapham DE
使用具有优异亲水性和可切换表面电荷的双功能材料深入分析血清中的糖蛋白唾液酸化
影响因子：
7.4
作者：
Dong, Xuefang;Qin, Hongqiang;Liang, Xinmiao
通讯作者：
Liang, Xinmiao