期刊
JOURNAL OF CHROMATOGRAPHY A
卷 1306, 期 -, 页码 20-26出版社
ELSEVIER SCIENCE BV
DOI: 10.1016/j.chroma.2013.07.060
关键词
Trypsin functionalized mesoporous silica nanotubes; Zirconia coated mesoporous silica nanotubes; Acombined procedure; Phosphopeptide enrichment
资金
- National Natural Science Foundation of China [20905039]
Trypsin functionalized mesoporous silica nanotubes bioreactor (TEMSN) and zirconia layer coated mesoporous silica nanotubes (ZrO2-MSN) were developed to deal with the long in-solution digestion time of phosphoprotein and detection difficulty of phosphorylated peptides, respectively. Trypsin was immobilized on the mesoporous silica nanotubes via epoxy group and TEMSN were used as a bioreactor for digestion of alpha-casein within 3 min. ZrO2-MSN were performed to enrich phosphopeptides selectively from in-solution digested peptide mixture of beta-casein to demonstrate that ZrO2-MSN possessed remarkable selectivity for phosphorylated peptides even at 100/1 molar ratio of BSA/beta-casein. The selective ability of ZrO2-MSN was also investigated in comparison to ZrO2 nanoparticles (ZrO2 NP). Moreover, phosphorylated peptides at the femtomole (2.5 fmol) level can also be detected with high S/N (signal-to-noise) ratio. Phosphopeptides enriched from TEMSN-bioreactor digested peptide mixture of alpha-casein was also performed to evaluate the cooperative performance of TEMSN and ZrO2-MSN platform. The experimental results indicated that TEMSN-bioreactor digestion changed the distribution of relative abundance of phosphopeptides and improved the relative intensity of partial phosphopeptides. This analytical strategy has also been applied to the identification of phosphopeptides isolated from non-fat bovine milk and got a comparable results compared with other materials cited from the literature. By matrix-assisted laser desorption/ionization mass spectrometry (MALDI-TOF MS), TEMSN and ZrO2-MSN were combined together for the rapid and comprehensive analysis of phosphoprotein. (C) 2013 Elsevier B.V. All rights reserved.
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