Chemical methods for glycoprotein discovery.
An important frontier in glycoproteomics is the discovery of proteins with post-translational glycan modifications. The first step in glycoprotein identification is the isolation of glycosylated proteins from the remainder of the proteome. New enzymatic and metabolic methods are being used to chemically tag proteins to enable their isolation. Once isolated, glycoproteins can be identified by mass spectrometry. Additional information can be obtained by using either enzymatic or chemoselective reactions to incorporate isotope labels at specific sites of glycosylation. Isotopic labeling facilitates mass spectrometry-based confirmation of glycoprotein identity, identification of glycosylation sites, and quantification of the extent of modification. By combining chemical tagging for isolation and isotope labeling for mass spectrometry analysis, researchers are developing highly effective strategies for glycoproteomics. These techniques are enabling cancer biologists to identify biomarkers whose glycosylation state correlates with disease states, and developmental biologists to characterize stage-specific changes in glycoprotein expression. Next-generation methods will make functional analyses of the glycoproteome possible, including the discovery of glycoprotein interaction partners and the identification of enzymes responsible for synthesis of particular glycan structures.
Bond
Michelle R
MR
Department of Chemistry, Stanford University, Keck Science Building, Stanford, CA 94305, USA.
Kohler
Jennifer J
JJ
D015415:Biomarkers / Q000032:analysis; D004798:Enzymes / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification* / Q000378:metabolism; D006031:Glycosylation; D007553:Isotope Labeling / Q000379:methods; D013058:Mass Spectrometry / Q000379:methods; D009369:Neoplasms / Q000378:metabolism / Q000473:pathology; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D040901:Proteomics / Q000379:methods
D015415:Biomarkers; D004798:Enzymes; D006023:Glycoproteins; D011134:Polysaccharides