Application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry with delayed ion extraction to ganglioside analyses

Sugiyama, Eiko; Hara, Atsushi; Uemura, Keiichi; Taketomi, Tamotsu

doi:10.1093/glycob/7.5.719

Cited by 62 publications

(55 citation statements)

References 32 publications

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“…The 2AB-labeled oligosaccharides fractionated by gel filtration HPLC on a Superdex peptide column were collected and individually analyzed by DE MALDI-TOF MS in the linear mode by Voyager-DE STR-H (Applied Biosystems, Foster City, CA) (21,22). Each sample (1 pmol) was mixed with a matrix, 2,5-dihydroxybenzoic acid, on a sample plate well, dried under an air stream, and then analyzed by MS in the negative or positive mode.…”

Section: Delayed Extraction Matrix-assisted Laser Desorption Ionizatimentioning

confidence: 99%

Identification of a Novel Chondroitin Hydrolase in Caenorhabditis elegans

Kaneiwa

Yamada

Mizumoto

et al. 2008

Journal of Biological Chemistry

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Section: Delayed Extraction Matrix-assisted Laser Desorption Ionizatimentioning

confidence: 99%

Identification of a Novel Chondroitin Hydrolase in Caenorhabditis elegans

Kaneiwa

Yamada

Mizumoto

et al. 2008

Journal of Biological Chemistry

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“…mass spectra in the positive ion mode of the isolated tetra-and hexasaccharide fractions were recorded in the linear mode in a Voyager DE-RP/Pro (PerSeptive Biosystems, Framingham, MA) (50). Each oligosaccharide fraction (30 pmol) was mixed with a small volume of an aqueous solution (10 mg/ml) of a matrix, 2,5-dihydroxybenzoic acid.…”

Section: Delayed Extraction Matrix-assisted Laser Desorption Ionizatimentioning

confidence: 99%

Specificities of Three Distinct Human Chondroitin/Dermatan N-Acetylgalactosamine 4-O-Sulfotransferases Demonstrated Using Partially Desulfated Dermatan Sulfate as an Acceptor

Mikami

Mizumoto

Kago

et al. 2003

Journal of Biological Chemistry

112

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4-O-Sulfation of GalNAc is a high frequency modification of chondroitin sulfate and dermatan sulfate (DS), and three major GalNAc 4-O-sulfotransferases including dermatan 4-O-sulfotransferase-1 (D4ST-1) and chondroitin 4-O-sulfotransferases-1 and -2 (C4ST-1 and -2) have been identified. 4-O-Sulfation of GalNAc during DS biosynthesis had long been postulated to be a prerequisite for iduronic acid (IdoUA) formation by C5-epimerization of GlcUA. This hypothesis has recently been argued based on enzymological studies using microsomes that C5-epimerization precedes 4-O-sulfation, which was further supported by the specificity of the cloned D4ST-1 with predominant preference for IdoUA-GalNAc flanked by GlcUA-GalNAc over IdoUA-GalNAc flanked by IdoUA-GalNAc in exhaustively desulfated dermatan. Whereas the counterproposal explains the initial reactions, apparently it cannot rationalize the synthetic mechanism of IdoUA-GalNAc(4-O-sulfate)-rich clusters typical of mature DS chains. In this study, we examined detailed specificities of the three recombinant human 4-O-sulfotransferases using partially desulfated DS as an acceptor. Enzymatic analysis of the transferase reaction products showed that D4ST-1 far more efficiently transferred sulfate to GalNAc residues in -IdoUA-GalNAc-IdoUA-than in -GlcUA-GalNAc-GlcUA-sequences. In contrast, C4ST-1 showed the opposite preference, and C4ST-2 used GalNAc residues in both sequences to comparable degrees, being consistent with its phylogenetic relations to D4ST-1 and C4ST-1. Structural analysis of the oligosaccharides, which were isolated after chondroitinase AC-I digestion of the 35 S-labeled transferase reaction products, revealed for the first time that D4ST-1, as compared with C4ST-1 and C4ST-2, most efficiently utilized GalNAc residues located not only in the sequence -IdoUA-GalNAc-IdoUA-but also in -GlcUA-GalNAc-IdoUA-and -IdoUA-GalNAc-GlcUA-. The isolated oligosaccharide structures also suggest that 4-O-sulfation promotes subsequent 4-O-sulfation of GalNAc in the neighboring disaccharide unit.

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“…MALDI-TOF mass spectra in the positive or negative ion mode of the 2AB-derivatized linkage hexasaccharide from C. elegans chondroitin were recorded on a Voyager DE-RP/Pro (PerSeptive Biosystems, Framingham, MA) in the linear mode (33). An aqueous solution of the 2AB-derivative of the linkage hexasaccharide, which was purified by gel filtration HPLC on an Asahipak GS320 column (32), was mixed with an equal volume of the aqueous solution (10 mg/ml) of a matrix, 2,5-dihydroxybenzoic acid.…”

Section: Delayed Extraction Matrix-assisted Laser Desorption Ionizatimentioning

confidence: 99%

Determination of the Glycosaminoglycan-Protein Linkage Region Oligosaccharide Structures of Proteoglycans from Drosophila melanogaster and Caenorhabditis elegans

Yamada

Okada

Ueno

et al. 2002

Journal of Biological Chemistry

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Caenorhabditis elegans and Drosophila melanogaster are relevant models for studying the roles of glycosaminoglycans (GAG) during the development of multicellular organisms. The genome projects of these organisms have revealed the existence of multiple genes related to GAG-synthesizing enzymes. Although the putative genes encoding the enzymes that synthesize the GAG-protein linkage region have also been identified, there is no direct evidence that the GAG chains bind covalently to core proteins. This study aimed to clarify whether GAG chains in these organisms are linked to core proteins through the conventional linkage region tetrasaccharide sequence found in vertebrates and whether modifications by phosphorylation and sulfation reported for vertebrates are present also in invertebrates. The linkage region oligosaccharides were isolated from C. elegans chondroitin in addition to D. melanogaster heparan and chondroitin sulfate after digestion with the respective bacterial eliminases and were then derivatized with a fluorophore 2-aminobenzamide. Their structures were characterized by gel filtration and anion-exchange high performance liquid chromatography in conjunction with enzymatic digestion and matrix-assisted laser desorption ionization time-of-flight spectrometry, which demonstrated a uniform linkage tetrasaccharide structure of -GlcUA-Gal-Gal-Xyl-or -GlcUA-Gal-Gal-Xyl(2-O-phosphate)-for C. elegans chondroitin and D. melanogaster CS, respectively. In contrast, the unmodified and phosphorylated counterparts were demonstrated in heparan sulfate of adult flies at a molar ratio of 73:27, and in that of the immortalized D. melanogaster S2 cell line at a molar ratio of 7:93, which suggests that the linkage region in the fruit fly first becomes phosphorylated uniformly on the Xyl residue and then dephosphorylated. It has been established here that GAG chains in both C. elegans and D. melanogaster are synthesized on the core protein through the ubiquitous linkage region tetrasaccharide sequence, suggesting that indispensable functions of the linkage region in the GAG synthesis have been well conserved during evolution. Mutations affecting the genes encoding putative proteins related to GAG biosynthetic enzymes have also been described for these organisms. Mutations in the tout velu (ttv) gene of D. melanogaster cause defects in Hedgehog movement in mosaic wing discs (1, 6). The ttv gene is a putative ortholog of vertebrate EXT1, which encodes a heparan polymerase and is associated with the hereditary multiple exostoses syndrome in humans (7). The pipe gene, which affects dorsal-ventral patterning of D. melanogaster development, encodes a homolog of vertebrate HS 2-O-sulfotransferase (2, 8). The sugarless and sulfateless genes, both of which affect the fibroblast growth factor signaling during the D. melanogaster development, en-

show abstract

Application of matrix-assisted laser desorption ionization time-of-flight mass spectrometry with delayed ion extraction to ganglioside analyses

Cited by 62 publications

References 32 publications

Identification of a Novel Chondroitin Hydrolase in Caenorhabditis elegans

Identification of a Novel Chondroitin Hydrolase in Caenorhabditis elegans

Specificities of Three Distinct Human Chondroitin/Dermatan N-Acetylgalactosamine 4-O-Sulfotransferases Demonstrated Using Partially Desulfated Dermatan Sulfate as an Acceptor

Determination of the Glycosaminoglycan-Protein Linkage Region Oligosaccharide Structures of Proteoglycans from Drosophila melanogaster and Caenorhabditis elegans

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