Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

Bowman, Michael J.; Zaia, Joseph

doi:10.1021/ac070581b

Cited by 96 publications

(118 citation statements)

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“…Recently, alternative isotopetagging methods using deuterium-labelled compounds, such as 2-aminobenzoic acid its derivatives, and permethylation, have been proposed by other groups. [22][23][24] All of these studies prove the utility of isotope-tagging methods for the quantitative analysis of glycosylation.…”

Section: Discussionmentioning

confidence: 87%

Alteration of N‐glycosylation in the kidney in a mouse model of systemic lupus erythematosus: relative quantification of N‐glycans using an isotope‐tagging method

et al. 2009

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Summary Changes in the glycan structures of some glycoproteins have been observed in autoimmune diseases such as systemic lupus erythematosus (SLE) and rheumatoid arthritis. A deficiency of α‐mannosidase II, which is associated with branching in N‐glycans, has been found to induce SLE‐like glomerular nephritis in a mouse model. These findings suggest that the alteration of the glycosylation has some link with the development of SLE. An analysis of glycan alteration in the disordered tissues in SLE may lead to the development of improved diagnostic methods and may help to clarify the carbohydrate‐related pathogenic mechanism of inflammation in SLE. In this study, a comprehensive and differential analysis of N‐glycans in kidneys from SLE‐model mice and control mice was performed by using the quantitative glycan profiling method that we have developed previously. In this method, a mixture of deuterium‐labelled N‐glycans from the kidneys of SLE‐model mice and non‐labelled N‐glycans from kidneys of control mice was analysed by liquid chromatography/mass spectrometry. It was revealed that the low‐molecular‐mass glycans with simple structures, including agalactobiantennary and paucimannose‐type oligosaccharides, markedly increased in the SLE‐model mouse. On the other hand, fucosylated and galactosylated complex type glycans with high branching were decreased in the SLE‐model mouse. These results suggest that the changes occurring in the N‐glycan synthesis pathway may cause the aberrant glycosylations on not only specific glycoproteins but also on most of the glycoproteins in the SLE‐model mouse. The changes in glycosylation might be involved in autoimmune pathogenesis in the model mouse kidney.

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Section: Discussionmentioning

confidence: 87%

Alteration of N‐glycosylation in the kidney in a mouse model of systemic lupus erythematosus: relative quantification of N‐glycans using an isotope‐tagging method

et al. 2009

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“…To circumvent these problems, LC/MS procedures have been developed that utilize linear equations to correlate the concentration of underivatized disaccharides with the detection of molecular standards (10,(17)(18)(19)(20). Another approach is to use mass-tagging techniques based on differentially isotopically labeled tags introduced into oligosaccharides derived from intact chains (16,(21)(22)(23)(24). Here we describe a mass-tagging technique that allows quantitative analysis of disaccharide composition and ratiometric comparisons between samples.…”

mentioning

confidence: 99%

Evolutionary Differences in Glycosaminoglycan Fine Structure Detected by Quantitative Glycan Reductive Isotope Labeling

Lawrence

Olson

Steele

et al. 2008

Journal of Biological Chemistry

174

195

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To facilitate qualitative and quantitative analysis of glycosaminoglycans, we tagged the reducing end of lyase-generated disaccharides with aniline-containing stable isotopes ( 12 C 6 and 13 C 6 ). Because different isotope tags have no effect on chromatographic retention times but can be discriminated by a mass detector, differentially isotope-tagged samples can be compared simultaneously by liquid chromatography/mass spectrometry and quantified by admixture with known amounts of standards. The technique is adaptable to all types of glycosaminoglycans, and its sensitivity is only limited by the type of mass spectrometer available. We validated the method using commercial heparin and keratan sulfate as well as heparan sulfate isolated from mutant and wild-type Chinese hamster ovary cells, and select tissues from mutant and wild-type mice. This new method provides more robust, reliable, and sensitive means of quantitative evaluation of glycosaminoglycan disaccharide compositions than existing techniques allowing us to compare the chondroitin and heparan sulfate compositions of Hydra vulgaris, Drosophila melanogaster, Caenorhabditis elegans, and mammalian cells. Our results demonstrate significant differences in glycosaminoglycan structure among these organisms that might represent evolutionarily distinct functional motifs.Metazoans make several types of sulfated glycosaminoglycans (GAGs), 2 including keratan sulfate (KS), chondroitin sulfate/dermatan sulfate (CS/DS), and heparan sulfate/heparin (HS). Each type of chain consists of unique disaccharide units. KS consists of galactose (Gal) and GlcNAc ([Gal␤1, 4GlcNAc␤1,3] n ) with variable sulfation at C6 of either sugar. CS/DS assembles as a copolymer of GlcA␤1,3GalNAc␤1,4 and then undergoes various processing reactions, including C5 epimerization of a portion of GlcA units to iduronic acid in DS, O-sulfation at C2 and more rarely at C3 of the uronic acids, and O-sulfation at C4 and C6 of the GalNAc residues (1). HS is the most highly modified GAG, consisting initially of GlcA␤1,4GlcNAc␣1,4 units, which then undergo variable processing by GlcNAc N-deacetylation and N-sulfation, C5 epimerization of some GlcA units to iduronic acid, and O-sulfate addition to C2 of the uronic acids and C6 and more rarely at C3 of the glucosamine units (2). The arrangement of the modified residues along the chain creates binding sites for numerous growth factors, enzymes, and extracellular matrix proteins. The structural variation that can occur makes sulfated GAG chains one of the most complex classes of macromolecules found in nature.GAG fine structure is typically assessed by analyzing the disaccharide composition of an isolated mixture of chains. A number of techniques have been developed to accomplish this task that rely on chemical or enzymatic depolymerization of the chains into their constituent disaccharides, followed by separation via anion exchange chromatography, reversed-phase chromatography with ion pairing agents, or capillary electrophoresis. These techniques separate...

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“…Alternative glycolabeling enrichment chemistries include the oxidation of cell surface polysaccharides on living cells combined with subsequent biocytin hydrazide labeling (30), but such approaches have been developed for cell culture and have not yet been applied in vivo. Several approaches have been developed that introduce a stable isotope to a given glycan via chemical derivatization (31,32). Glycans are typically derivatized prior to analysis either by tagging the reducing terminus with a chromophore when subsequent analyses are chromatographic or by permethylation when the sample is to be analyzed by MS. A typical work flow for these in vitro labeling approaches involves the parallel release of glycans from the sample populations under investigation and derivatization with an isotopic label after which the samples are mixed and then analyzed by MS.…”

Section: Technical Challenges Associated With Membrane Proteomicsmentioning

confidence: 99%

Overcoming Key Technological Challenges in Using Mass Spectrometry for Mapping Cell Surfaces in Tissues

Griffin

Schnitzer

2011

Molecular & Cellular Proteomics

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Plasma membranes form a critical biological interface between the inside of every cell and its external environment. Their roles in multiple key cellular functions make them important drug targets. However the protein composition of plasma membranes in general is poorly defined as the inherent properties of lipid embedded proteins, such as their hydrophobicity, low abundance, poor solubility and resistance to digestion and extraction makes them difficult to isolate, solubilize, and identify on a large scale by traditional mass spectrometry methods. Here we describe some of the significant advances that have occurred over the past ten years to address these challenges including: i) the development of new and improved membrane isolation techniques via either subfractionation or direct labeling and isolation of plasma membranes from cells and tissues; ii) modification of mass spectrometry methods to adapt to the hydrophobic nature of membrane proteins and peptides; iii) improvements to digestion protocols to compensate for the shortage of trypsin cleavage sites in lipid-embedded proteins, particularly multi-spanning proteins, and iv) the development of numerous bioinformatics tools which allow not only the identification and quantification of proteins, but also the prediction of membrane protein topology, membrane post-translational modifications and subcellular localization. This review emphasis the importance and difficulty of defining cells in proper patho-and physiological context to maintain the in vivo reality. We focus on how key technological challenges associated with the isolation and identification of cell surface proteins in tissues using mass spectrometry are being addressed in order to identify and quantify a comprehensive plasma membrane for drug and target discovery efforts. Molecular & Cellular

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Tags for the Stable Isotopic Labeling of Carbohydrates and Quantitative Analysis by Mass Spectrometry

Cited by 96 publications

References 35 publications

Alteration of N‐glycosylation in the kidney in a mouse model of systemic lupus erythematosus: relative quantification of N‐glycans using an isotope‐tagging method

Alteration of N‐glycosylation in the kidney in a mouse model of systemic lupus erythematosus: relative quantification of N‐glycans using an isotope‐tagging method

Evolutionary Differences in Glycosaminoglycan Fine Structure Detected by Quantitative Glycan Reductive Isotope Labeling

Overcoming Key Technological Challenges in Using Mass Spectrometry for Mapping Cell Surfaces in Tissues

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