Exploring the glycoproteomics landscape with advanced MS technologies

Abstract

The advance of glycoproteomic technologies has offered unique insights into the importance of glycosylation in determining the functional roles of a protein within a cell. Biologically active glycoproteins include the categories of enzymes, hormones, proteins involved in cell proliferation, cell membrane proteins involved in cell-cell recognition, and communication events or secreted proteins, just to name a few. The recent progress in analytical instrumentation, methodologies, and computational approaches has enabled a detailed exploration of glycan structure, connectivity, and heterogeneity, underscoring the staggering complexity of the glycome repertoire in a cell. A variety of approaches involving the use of spectroscopy, MS, separation, microfluidic, and microarray technologies have been used alone or in combination to tackle the glycoproteome challenge, the research results of these efforts being captured in an overwhelming number of annual publications. This work is aimed at reviewing the major developments and accomplishments in the field of glycoproteomics, with focus on the most recent advancements (2012-2014) that involve the use of capillary separations and MS detection.

Electron-Transfer/Higher-Energy Collision Dissociation (EThcD)-Enabled Intact Glycopeptide/Glycoproteome Characterization

“…However, due to its heterogeneous nature, fragmenting an intact glycopeptide has always been difficult and yet no single fragmentation technique is able to generate a complete picture in a single MS/MS spectrum [6, 7, 14, 15]. Peaks resulting from glycosidic bond cleavages dominate spectra generated by collision-induced dissociation (CID) with little knowledge of glycosylation sites and amino acid sequences, whereas c/z-ion series in ETD type of experiments yield the glycosylation site and peptide identity with little information on glycan side chain composition [16, 17].…”

The Fine Art of Destruction: A Guide to In‐Depth Glycoproteomic Analyses—Exploiting the Diagnostic Potential of Fragment Ions

“…Here, different liquid chromatography (LC) and solid phase extraction (SPE)–based methods can be employed, such as hydrophilic interaction liquid chromatography (HILIC) . Arguably, the biggest leap in the field of glycoproteomics can be attributed to advances in mass spectrometry instrumentation and software development . The advent of high‐resolution mass spectrometers, including orbitrap instruments and next‐generation Q‐TOF instruments, along with new and complementary fragmentation techniques, namely higher energy collision dissociation (HCD) and electron transfer dissociation (ETD), has tremendously increased the throughput, depth and confidence of glycoproteomic analyses.…”

Recent advances in mass spectrometric analysis of glycoproteins

“…Over the past few years, several published articles have reviewed the analytical methods developed for the characterization of glycans and glycoproteins with focus on chromatography, electrophoresis, electrochemistry, and MS [, ]. This review specifically summarizes and highlights recent developments (2014–2016) in the fields of glycomics and glycoproteomics with emphasis on the latest advancements involving MS imaging (MSI), ion mobility (IM) MS, and capillary separations.…”