Mesoporous zirconium oxide nanomaterials effectively enrich phosphopeptides for mass spectrometry-based phosphoproteomics

Abstract

This work represents the first use of mesoporous zirconium oxide nanomaterials for highly effective and selective enrichment of phosphorylated peptides.Reversible protein phosphorylation is a ubiquitous post-translational modification that plays a vital role in the control of many biological processes such as cellular growth, division, and signaling. 1 Aberrant phosphorylation is known to be one of the underlying mechanisms for many human diseases, most notably cancer. 1a,c Mass spectrometry (MS) has become the most important and powerful tool for the analysis of protein phosphorylation due to its sensitivity, speed, simplicity, separation, and specificity. 2 While MS techniques have been successfully applied to determine the phosphorylation state of a single protein/peptide, MS analysis of phosphorylation on a proteome-wide scale still poses substantial challenges due to the low abundance of phosphoproteins and substoichiometric phosphorylation. 3 Therefore, isolation and enrichment of the phosphoproteins/peptides are essential for MS-based phosphoproteomics. 4 The affinity based method such as immobilized metal ion affinity chromatography (IMAC) 5 using Ga(III), Fe(III), or other metals, has been widely used for phosphopeptides enrichment. Recently microparticles of titanium dioxide (TiO 2 ), 6a,b zirconium oxide (ZrO 2 ), 6c and other metal oxides 6d-e have demonstrated higher specificity for trapping phosphate than the conventional IMAC beads since such oxides rely on specific and reversible chemisorption of phosphate groups on their amphoteric surface and have less nonspecific binding. Additionally, nanoparticles, such as ZrO 2 , TiO 2 , Fe 2 O 3 , and titania-coated magnetic iron oxide (Fe 3 O 4 @TiO 2 ) nanoparticles, have recently been explored due to their potential higher capacities than the microparticles. 7 Mesoporous materials are nanostructured materials with pore sizes typically between 2−50 nm. 8 They have extremely large surface areas and have been utilized in many applications such as catalyst support and filtration. Such large surface areas, together with the many active surface sites, can be translated into even higher loading capacity for binding phosphate groups than micro-and nanoparticles. 9 In addition to their well-ordered nanoscale porous structures and © The Royal Society of Chemistry [year] Correspondence to: Song Jin, jin@chem.wisc.edu; Ying Ge, yge@physiology.wisc.edu. NIH Public Access Author ManuscriptChem Commun (Camb) flow-through capacity, they are chemically stable and can be easily prepared at reasonable cost. All these attributes would make them ideal for applications in MS-based phosphoroproteomics. Herein, we report the utility of ZrO 2 nanomaterials for simple and efficient enrichment of phosphopeptides with high specificity. This is the first application of mesoporous ZrO 2 nanomaterials for phosphopeptide enrichment.We chose ZrO 2 metal oxide because of its known amphoteric surface properties, 10 which facilitates preferable and reversable binding and r...

Zirconium oxide aerogel for effective enrichment of phosphopeptides with high binding capacity

“…It can be seen that the zirconia aerogel is highly porous, with cauliflower-type surface. By N 2 adsorption isotherms analysis [30], the average pore size is 10.2 nm, and the specific surface area is 416.0 m 2 g −1 , nearly tenfold higher than that of ZrO 2 nanoparticles [13] and five folds higher than that of the latest reported mesoporous ZrO 2 nanomaterials [16]. Such super-large surface area could offer enough active binding site and high binding capacity for the enrichment of phosphopeptides.…”

“…Furthermore, to perform the biological activity study instead of the qualitative analysis of phosphopeptides, the large-scale enrichment of phosphopeptides from samples could be easily achieve by materials with high binding capacity. Therefore, the preparation of material with large binding capacity and good selectivity for phosphopeptides is greatly required, and some novel materials, such as Fe 3 O 4 /TiO 2 cores/shell nanoparticles [12], ZrO 2 nanoparticles [13], mesoporous silica particles modified with TiO 2 [14,15], and mesoporous ZrO 2 nanomaterials [16] have been prepared. However, for metal oxides with further increased specific surface area, such as aerogels, the binding capacity for phosphopeptides can be improved obviously.…”

Mesoporous TiO2 aerogel for selective enrichment of phosphopeptides in rat liver mitochondria

“…Besides, higher sensitivity for phosphopeptides detection can be achieved by immobilized mesoporous metal oxides with high binding capacity on the MALDI plate [21,27,28]. However, the surface area of pure mesoporous MOAC materials is often less than 100 m 2 g −1 [24,29], and only that for those supported by silica matrices can get larger than 100 m 2 g −1 , but with complex synthesis procedure [22,23,30]. The limited specific surface area of the reported MOAC materials results in low binding capacity and low sensitivity for phosphopeptide detection.…”

Post-Genomics Nanotechnology Is Gaining Momentum: Nanoproteomics and Applications in Life Sciences

“…The field of nanotechnology has been associated with several proteomics applications such as phosphoproteomics/metal oxide nanoparticles, nanostructure surfaces for protein separation, and analytical detection of biomarker proteins using arrays techniques (Leitner, 2010;Nelson et al, 2009;Northen et al, 2007;Rissin et al, 2010). This merging between nanotechnology and proteomics has generated nanoproteomics, which is defined as a discipline of science involving the application of proteomics techniques aided by nanotechnology to enhance probing and evaluating protein systems (Archakov, 2007).…”