New Technologies for Glycomic Analysis: Toward a Systematic Understanding of the Glycome

Abstract

Carbohydrates are the most difficult class of biological molecules to study by high-throughput methods owing to the chemical similarities between the constituent monosaccharide building blocks, template-less biosynthesis, and the lack of clearly identifiable consensus sequences for the glycan modification of cohorts of glycoproteins. These molecules are crucial for a wide variety of cellular processes ranging from cell-cell communication to immunity, and they are altered in disease states such as cancer and inflammation. Thus, there has been a dedicated effort to develop glycan analysis into a high-throughput analytical field termed glycomics. Herein we highlight major advances in applying separation, mass spectrometry, and microarray methods to the fields of glycomics and glycoproteomics. These new analytical techniques are rapidly advancing our understanding of the importance of glycosylation in biology and disease.

Mapping posttranscriptional regulation of the human glycome uncovers microRNA defining the glycocode

“…Lectin microarrays, in which carbohydrate-binding proteins are probes for glycan structure, provide a systems-level view of the glycome (Fig. 1A) (18)(19)(20). These microarrays give specific information on the repertoire of glycans present, e.g., high mannose epitopes, branching patterns, and terminal α-2,3-or α-2,6-sialic acids, in a high-throughput format.…”

“…These microarrays give specific information on the repertoire of glycans present, e.g., high mannose epitopes, branching patterns, and terminal α-2,3-or α-2,6-sialic acids, in a high-throughput format. Unlike other glycomic methods, both the N-and O-linked glycome are observed (19), and the data are in the same format as mRNA and miRNA datasets. We analyzed the NCI-60, a cancer cell line panel containing 59 cell lines from nine tissue types, using our lectin microarray technology.…”

Advances in purification and separation of posttranslationally modified proteins

“…5, H). These PTMs play important roles in many key biological processes, including protein folding, signaling, enzymatic activity, molecular trafficking, cell adhesion and protein turnover [163,164]. Glycosylation is believed to be more prevalent than phosphorylation and it is estimated that at least 50 % of all proteins are modified by carbohydrates [163].…”

“…In further contrast to N-glycosylation, there is no known conserved sequence motif for O-glycosylation [1,164]. C-glycosylation is rare, and has only been found at tryptophan residues, the C2 position of the indole ring being a mannosylation target [163]. Non-enzymatic protein glycation may occur at lysine or arginine amino groups, or the N-terminal amino acid residues of proteins.…”

Can Microfluidics boost the Map of Glycome Code?

“…The term "glycome" describes the complete repertoire of glycans and glycoconjugates that cells produce under specified conditions of time, space, and environment [1][2][3][4]. "Glycomics" refers to studies that profile the glycome [5,6].…”