For the first time, dual metal ions (Ti4+-Zr4+) were successfully modified into the channel of magnetic mesoporous silica to obtain an affinity probe for highly selective capture of endogenous phosphopeptides in biological samples. The newly prepared Fe3O4@mSiO2@Ti4+-Zr4+ composites possessed the advantages of ordered mesoporous channels, superparamagnetism, and enhanced affinity properties of dual metal ions of Ti4+ and Zr4+. The phosphopeptide enrichment efficiency of the Fe3O4@mSiO2@Ti4+-Zr4+ composite was investigated, and the result indicated an ultrahigh size exclusive ability (weight ratio of β-casein tryptic digests, BSA, and α-casein protein reached up to 1:1000:1000). Compared to magnetic affinity probes with single metal ions (Fe3O4@mSiO2@Ti4+, Fe3O4@mSiO2@Zr4+), the composite possessed stronger specificity, higher sensitivity, and better efficiency; and more importantly, it showed much enhanced enrichment ability towards both mono- and multi-phosphorylated peptides. Additionally, by utilizing the Fe3O4@mSiO2@Ti4+-Zr4+ affinity probe, a total number of 104 endogenous phosphopeptides including 95 mono-phosphopeptides and 9 multi-phosphopeptides were captured and identified from human saliva, indicating the great potential for the application of the novel probe for the peptidome analysis in the future.
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La Barbera G, Capriotti AL, Cavaliere C, Ferraris F, Laus M, Piovesana S, Sparnacci K, Laganà A (2018) Development of an enrichment method for endogenous phosphopeptide characterization in human serum. Anal Bioanal Chem 410:1177–1185
Marshall J, Kupchak P, Zhu W, Yantha J, Vrees T, Furesz S, Jacks K, Smith C, Kireeva I, Zhang R, Takahashi M, Stanton E, Jackowski G (2003) Processing of serum proteins underlies the mass spectral fingerprinting of myocardial infarction. J Proteome Res 2:361–372
Dallas DC, Guerrero A, Parker EA, Robinson RC, Gan J, German JB, Barile D, Lebrilla CB (2015) Current peptidomics: applications, purification, identification, quantification, and functional analysis. Proteomics 15:1026–1038
Liu Q, Sun N, Gao M, Deng C-h (2018) Magnetic binary metal–organic framework as a novel affinity probe for highly selective capture of endogenous phosphopeptides. ACS Sustain Chem Eng 6:4382–4389
Beltran L, Cutillas PR (2012) Advances in phosphopeptide enrichment techniques for phosphoproteomics. Amino Acids 43:1009–1024
Su J, He X, Chen L, Zhang Y (2018) Adenosine phosphate functionalized magnetic mesoporous graphene oxide nanocomposite for highly selective enrichment of phosphopeptides. ACS Sustain Chem Eng 6:2188–2196
Boersema PJ, Mohammed S, Heck AJR (2009) Phosphopeptide fragmentation and analysis by mass spectrometry. J Mass Spectrom 44:861–878
Lin H, Chen H, Shao X, Deng C (2018) A capillary column packed with a zirconium(IV)-based organic framework for enrichment of endogenous phosphopeptides. Microchim Acta 185:562
Li Y, Liu L, Wu H, Deng C (2019) Magnetic mesoporous silica nanocomposites with binary metal oxides core-shell structure for the selective enrichment of endogenous phosphopeptides from human saliva. Anal Chim Acta 1079:111–119
Villanueva J, Philip J, Entenberg D, Chaparro CA, Tanwar MK, Holland EC, Tempst P (2004) Serum peptide profiling by magnetic particle-assisted, automated sample processing and MALDI-TOF mass spectrometry. Anal Chem 76:1560–1570
Lan X, Liao D, Wu S, Wang F, Sun J, Tong Z (2015) Rapid purification and characterization of angiotensin converting enzyme inhibitory peptides from lizard fish protein hydrolysates with magnetic affinity separation. Food Chem 182:136–142
Yao J, Sun N, Deng C, Zhang X (2015) Designed synthesis of graphene @titania @mesoporous silica hybrid material as size-exclusive metal oxide affinity chromatography platform for selective enrichment of endogenous phosphopeptides. Talanta 150:296–301
Yan Y, Zhang X, Deng C (2014) Designed synthesis of titania nanoparticles coated hierarchically ordered macro/mesoporous silica for selective enrichment of phosphopeptides. ACS Appl Mater Interfaces 6:5467–5471
Chen CT, Chen YC (2005) Fe3O4/TiO2 Core/shell nanoparticles as affinity probes for the analysis of phosphopeptides using TiO2 surface-assisted laser desorption/ionization mass spectrometry. Anal Chem 77:5912–5919
Yan Y, Zheng Z, Deng C, Zhang X, Yang P (2013) Facile synthesis of Ti4+−immobilized Fe3O4@polydopamine core-shell microspheres for highly selective enrichment of phosphopeptides. Chem Commun 49:5055–5057
Qi D, Mao Y, Lu J, Deng C, Zhang X (2010) Phosphate-functionalized magnetic microspheres for immobilization of Zr4+ ions for selective enrichment of the phosphopeptides. J Chromatogr A 1217:2606–2617
Andersson L, Porath J (1986) Isolation of phosphoproteins by immobilized metal (Fe3+) affinity chromatography. Anal Biochem 154:250–254
Kinoshita E, Kinoshita-Kikuta E, Takiyama K, Koike T (2006) Phosphate-binding tag, a new tool to visualize phosphorylated proteins. Mol Cell Proteomics 5:749–757
Hu Q, Hu S, Zhang Z, Zhou X, Yang S, Zhang Y, Chen X (2011) Fe3+−immobilized nanoparticle-modified capillary for capillary electrophoretic separation of phosphoproteins and non-phosphoproteins. Electrophoresis 32:2867–2873
Posewitz MC, Tempst P (1999) Immobilized gallium(III) affinity chromatography of phosphopeptides. Anal Chem 71:2883–2892
Ficarro SB, Adelmant G, Tomar MN, Zhang Y, Cheng VJ, Marto JA (2009) Magnetic bead processor for rapid evaluation and optimization of parameters for phosphopeptide enrichment. Anal Chem 81:4566–4575
Yu Z, Han G, Sun S, Jiang X, Chen R, Wang F, Wu R, Ye M, Zou H (2009) Preparation of monodisperse immobilized Ti4+ affinity chromatography microspheres for specific enrichment of phosphopeptides. Anal Chim Acta 636:34–41
Han G, Ye M, Zou H (2008) Development of phosphopeptide enrichment techniques for phosphoproteome analysis. Analyst. 133:1128–1138
Lai ACY, Tsai CF, Hsu CC, Sun YN, Chen YJ (2012) Complementary Fe3+- and Ti4+-immobilized metal ion affinity chromatography for purification of acidic and basic phosphopeptides. Rapid Commun Mass Sp 26:2186–2194
Sun N, Deng C, Li Y, Zhang X (2014) Size-exclusive magnetic graphene/mesoporous silica composites with titanium(IV)-immobilized pore walls for selective enrichment of endogenous phosphorylated peptides. ACS Appl Mater Interfaces 6:11799–11804
Yin P, Wang Y, Li Y, Deng C, Zhang X, Yang P (2012) Preparation of sandwich-structured graphene/mesoporous silica composites with C8-modified pore wall for highly efficient selective enrichment of endogenous peptides for mass spectrometry analysis. Proteomics 12:2784–2791
Zhu GT, Li X-S, Gao Q, Zhao N-W, Yuan B-F, Feng Y-Q (2012) Pseudomorphic synthesis of monodisperse magnetic mesoporous silica microspheres for selective enrichment of endogenous peptides. J Chromatogr A 1224:11–18
Tian R, Zhang H, Ye M, Jiang X, Hu L, Li X, Bao X, Zou H (2010) Selective extraction of peptides from human plasma by highly ordered mesoporous silica particles for peptidome analysis. Angew Chem Int Ed 46:962–965
Zhang Q, Xiong Z, Wan H, Chen X, Zou H (2016) Facile preparation of mesoporous carbon-silica-coated graphene for the selective enrichment of endogenous peptides. Talanta 146:272–278
Long X-y, Li J-y, Sheng D, Lian H-Z (2016) Low-cost iron oxide magnetic nanoclusters affinity probe for the enrichment of endogenous phosphopeptides in human saliva. RSC Adv 6:96210–96222
Kweon HK, Håkansson K (2006) Selective zirconium dioxide-based enrichment of phosphorylated peptides for mass spectrometric analysis. Anal Chem 78:1743–1749
Yao J, Sun N, Wang J, Xie Y, Deng C, Zhang X (2017) Rapid synthesis of titanium(IV)-immobilized magnetic mesoporous silica nanoparticles for endogenous phosphopeptides enrichment. Proteomics 17:1600320
Wu Y, Liu Q, Xie Y, Deng C (2018) Core-shell structured magnetic metal-organic framework composites for highly selective enrichment of endogenous N-linked glycopeptides and phosphopeptides. Talanta 190:298–312
Bennick A (1982) Salivary proline-rich proteins. Mol Cell Biochem 45:83–99
Robinovitch MR, Ashley RL, Iversen JM, Vigoren EM, Lamkin M (2001) Parotid salivary basic proline-rich proteins inhibit HIV-I infectivity. Oral Dis 7:86–93
This work was financially supported by the National Natural Science Foundation of China (21425518, 21405022, and 21675034) and National Basic Research Priorities Program of China (2013CB911201).
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Hu, X., Li, Y., Miao, A. et al. Dual metal cations coated magnetic mesoporous silica probe for highly selective capture of endogenous phosphopeptides in biological samples. Microchim Acta 187, 400 (2020). https://doi.org/10.1007/s00604-020-04323-6
- Endogenous phosphopeptide
- Molecular recognition
- Magnetic mesoporous probe
- Dual metal cations affinity