Advances in proteomics sample preparation and enrichment for phosphorylation and glycosylation analysis

Abstract

As the common and significant chemical modifications, post‐translational modifications (PTMs) play a key role in the functional proteome. Affected by the signal interference, low concentration, and insufficient ionization efficiency of impurities, the direct detection of PTMs by mass spectrometry (MS) still faces many challenges. Therefore, sample preparation and enrichment are an indispensable link before MS analysis of PTMs in proteomics. The rapid development of functionalized materials with diverse morphologies and compositions provides an avenue for sample preparation and enrichment for PTMs analysis. In this review, we summarize recent advances in the application of novel functionalized materials in sample preparation for phosphoproteomes and glycoproteomes analysis. In addition, this review specifically discusses the design and preparation of functionalized materials based on different enrichment mechanisms, and proposes research directions and potential challenges for proteomic PTMs research.

Innovative Chemical Tools to Address Analytical Challenges of Protein Phosphorylation and Glycosylation

“…Besides, Ye’s group has reported an affinity reagent derived from the Src homology 2 (SH2) domain as an alternative to antibodies to carry out the high-efficiency pTyr enrichment . Deng’s group has developed a series of functionalized magnetic nanomaterials to perform phosphorylation enrichment . Nevertheless, conventional affinity strategies are often faced with the problem that enriched targets are difficult to dissociate from the material surface due to the strong chelation or multivalent interaction with multiple phosphoryl groups in the case of the enrichment of multiply phosphorylated peptides (MPPs).…”

“…13 Deng's group has developed a series of functionalized magnetic nanomaterials to perform phosphorylation enrichment. 14 Nevertheless, conventional affinity strategies are often faced with the problem that enriched targets are difficult to dissociate from the material surface due to the strong chelation or multivalent interaction with multiple phosphoryl groups in the case of the enrichment of multiply phosphorylated peptides (MPPs). Therefore, a large amount of multisite phosphorylation information was not accessible.…”

Quartz Crystal Microbalance as a Holistic Detector for Quantifying Complex Organic Matrices during Liquid Chromatography: 1. Coupling, Characterization, and Validation

“…A challenge in trace analysis of targeted organic compounds is highly complex samples, such as environmental, 1,2 biological, 3,4 or food samples. 5−7 This is the case because abundant organic and inorganic constituents of the complex sample, other than the target analyte(s) and also known as the matrix, can have a variety of adverse effects on the analytical mode of detection.…”

Self-assembly of hydrazide-linked porous organic polymers rich in titanium for efficient enrichment of glycopeptides and phosphopeptides from human serum

“…Glycosylation and phosphorylation are two of the most common post-translational modifications (PTMs) of proteins. 1 PTMs regulate biological processes by altering various functions and activities of proteins such as cell growth, proliferation, differentiation, apoptosis and cell signaling. 2 Abnormal glycosylation and phosphorylation are popularly applied in clinical proteomics studies and biomarker discovery related to diseases.…”