Matrix-assisted laser desorption/ionization tandem mass spectrometry and post-source decay fragmentation study of phenylhydrazones of <i>N</i>-linked oligosaccharides from ovalbumin

Lattová, Erika; Perreault, Hélène; Krokhin, Oleg V.

doi:10.1016/j.jasms.2004.01.012

Cited by 43 publications

(55 citation statements)

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“…In our previous recent articles we introduced development phenylhydrazine (PHN) derivatization for labeling oligosaccharides from commercial sources [29,32]. The present study for the first time demonstrates application of this method for mass spectrometric analyses of carbohydrates from real complex biological material.…”

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confidence: 75%

“…Then, before each spot dried completely, PHN solution was spotted (0.5 L; 3 L of PHN reagent diluted in 12 L of deionized water and 3 L of ACN) and samples were left to dry at 37°C (ϳ30 min). MS/MS spectra of oligosaccharides were interpreted manually and detected structures of common N-glycan were compared with compositions described in previous studies [31][32][33]. For the assignment of all oligosaccharide fragment ions, the general nomenclature established by Domon and Costello was followed [34].…”

Section: Maldi-mass Spectrometric Analysismentioning

confidence: 99%

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Mass spectrometric profiling of N-linked oligosaccharides and uncommon glycoform in mouse serum with head and neck tumor

Lattová

Varma

Bezabeh

et al. 2008

J. Am. Soc. Mass Spectrom.

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N-linked oligosaccharides obtained from total serum of mice with implanted head and neck tumors were analyzed and compared with those from control samples of healthy mice. Methods used include a combination of a derivatization procedure with phenylhydrazine (PHN) and analysis by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). Oligosaccharides were enzymatically released from total serum with PNGaseF and purified by high-performance liquid chromatography (HPLC) on a reversed-phase column. Mass spectra contained ion peaks of labeled oligosaccharides and MS/MS experiments provided useful data for the structural elucidation of these compounds. More than 40 N-glycans with compositions characteristic of high-mannose, hybrid, complex, neutral, and sialylated structures were identified in the serum of tumoral mice. Significant differences between samples were observed with respect to the abundances of high mannose and hybrid glycans. These oligosaccharides showed higher relative intensities in the spectra obtained from the cancer sera. Complex sialylated oligosaccharides had similar abundances in both types of sera, with the exception of fucosylated biantennary disialylated oligosaccharide, which was mostly detected with lower abundance in control samples. In the MALDI spectra, several minor species corresponded to uncommon carbohydrates. These structures have been investigated in detail by MS/MS. Among these novel glycoforms, a few sialylated oligosaccharides without a free reducing end were identified. Also, glycans with an extra 60 u were observed and likely feature the presence of a 2-acetamido-2-deoxyoctose residue attached on antennae of 3-or 6-linked mannose. . It has been recognized that altered glycosylation is an universal feature of living cells and several studies have demonstrated that the changes in cellular glycosylation profiles can be also related to aggressive cancer cell behavior, such as tumor cell invasion and metastasis [2][3][4][5][6][7][8]. As cancer cells can produce oligosaccharides that are significantly different from those in normal cells, comparative studies between glycoprotein carbohydrates produced by malignant cells and normal cells may provide useful information for diagnosis, prognosis, and immunotherapy of tumors [9].Mass spectrometry (MS) has been used extensively as the most sensitive method for the analysis of carbohydrate molecules [10 -14]. Moreover, modern MS instruments allow the measurement of unseparated mixtures, and make possible the identification of trace amounts of glycans in biological materials [15]. Nonetheless, biochemical analysis of carbohydrates is generally complicated, not only because of their lower concentration relative to with proteins, but also because of the structural diversity of molecules, anomericity, linkage positions, and branching patterns of monosaccharides. Moreover, carboxylic groups cause sialylated oligosaccharides to ionize with less efficiency than neutral oligosaccharides, making MS analysis of glycans more difficu...

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confidence: 75%

Section: Maldi-mass Spectrometric Analysismentioning

confidence: 99%

Mass spectrometric profiling of N-linked oligosaccharides and uncommon glycoform in mouse serum with head and neck tumor

Lattová

Varma

Bezabeh

et al. 2008

J. Am. Soc. Mass Spectrom.

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“…The low-energy CID spectrum displayed in Figure 2a was generated in the linear ion-trap instrument for the (GlcNAc) 6 (Man) 3 glycan structure (m/z 2153). It was previously shown that this structure consists of three isomeric glycan structures (denoted in different colors) depicted as I, II, and III in the inset of Figure 2a [62,63]. The most abundant product ion observed in the spectrum is m/z 1894, corresponding to the facile loss of GlcNAc (Nacetyl glucosamine) residue from the precursor.…”

Section: (A) (A) (B) (C) (D)mentioning

confidence: 90%

Identification of isomeric N-glycan structures by mass spectrometry with 157 nm laser-induced photofragmentation

Devakumar

Mechref

Kang

et al. 2008

J. Am. Soc. Mass Spectrom.

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Characterization of structural isomers has become increasingly important and extremely challenging in glycobiology. This communication demonstrates the capability of ion-trap mass spectrometry in conjunction with 157 nm photofragmentation to identify different structural isomers of permethylated N-glycans derived from ovalbumin without chromatographic separation. The results are compared with collision-induced dissociation (CID) experiments. Photodissociation generates extensive cross-ring fragment ions as well as diagnostic glycosidic product ions that are not usually observed in CID MS/MS experiments. The detection of these product ions aids in characterizing indigenous glycan isomers. The ion trap facilitates MS n experiments on the diagnostic glycosidic fragments and cross-ring product ions generated through photofragmentation, thus allowing unambiguous assignment of all of the isomeric structures associated with the model glycoprotein used in this study. Photofragmentation is demonstrated to be a powerful technique for the structural characterization of glycans. , and protein regulations and interactions [6], is widely acknowledged [7]. To improve our understanding of these processes, it is important to explore glycan structure-function relationships. However, the detailed characterization of a glycan structure and its attributes remains a difficult task, given the microheterogeneity and diversity of these molecules. Importantly, the discrimination of numerous structural isomers differing in sequence, linkage, position, or branching features remains a challenging frontier of glycobiology.For years, the characterization of glycan structures has almost exclusively been accomplished by tandem mass spectrometry (MS/MS) [1,8,9] because of its high sensitivity and minimum sample requirements relative to nuclear magnetic resonance (NMR). Different combinations of ionization techniques, ion activation, and A glycan ion generally fragments in two ways: (1) glycosidic cleavages resulting from a bond rupture between two adjacent sugar residues and (2) cross-ring cleavages in which any two bonds on the same sugar unit are broken. Cross-ring fragment ions are commonly observed in high-energy collision-induced dissociation (CID) methods as demonstrated in the tandem TOF/TOF approach [34 -38]. A limitation of this technique, however, is its inability to perform multistage tandem mass spectrometry experiments. On the other hand, glycosidic cleavage ions are predominantly observed in low-energy activation methods and are mainly used to derive sequence and limited branching information. Most recently, demonstrated the use of low-energy activation to identify the structural isomers of glycans in complex mixtures with the help of sequential tandem mass spectrometry (MS n ). However, a disadvantage of CID is the decrease in both the degree and efficiency of dissociation with increasing mass and MS n events. Alternatively, other activation techniques have been used for Address reprint requests to Prof. James P.

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“…Symbols are the same as in Figure 1. companied by the following loss of water at m/z 694.2 45 could be assigned to the isomer with a branched antenna on 3-linked core mannose (1).…”

Section: Maldi-ms Analysis Of N-glycans From Mcf-7 and Cemmentioning

confidence: 99%

N-Glycomic Changes in Human Breast Carcinoma MCF-7 and T-Lymphoblastoid Cells After Treatment with Herceptin and Herceptin/Lipoplex

et al. 2010

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The humanized monoclonal antibody IgG1 in combination with chemotherapy has been demonstrated to enhance survival benefit in cancer treatment. Despite positive outcomes, some cancer cells develop multidrug resistance. Numerous mechanisms in cancers can be involved in the process of treatment therapy and most of them are not still well understood. To address how the carbohydrate moieties of cells are affected during treatment, the glycan profiles from the two most common cancer cell lines s human breast MCF-7 carcinoma and T-lymphoblastoid CEM cells s were studied here and compared with profiles after treatment with Herceptin alone or in combination with Lipofectamine mixed with plasmid DNA to form Lipoplex. N-Glycans were released from total cells by digestion with PNGaseF and analyzed by matrix-assisted laser desorption ionization mass spectrometry (MALDI-MS). In summary, both original cell lines showed a dominant occurrence of high-mannose glycans. After treatment, these structures were suppressed and biantennary core-fucosylated glycans originating from IgG1 were the major carbohydrate products identified in cells. The high incidence of additional fucosylated or nonfucosylated galactosylated oligosaccharides, which were not detected in original cells or Herceptin, varied with conditions and time of exposure of cells to the antibody. The results presented in this study provide strong evidence for a role of glycosylation during antibody treatment.

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Matrix-assisted laser desorption/ionization tandem mass spectrometry and post-source decay fragmentation study of phenylhydrazones of N-linked oligosaccharides from ovalbumin

Cited by 43 publications

References 40 publications

Mass spectrometric profiling of N-linked oligosaccharides and uncommon glycoform in mouse serum with head and neck tumor

Mass spectrometric profiling of N-linked oligosaccharides and uncommon glycoform in mouse serum with head and neck tumor

Identification of isomeric N-glycan structures by mass spectrometry with 157 nm laser-induced photofragmentation

N-Glycomic Changes in Human Breast Carcinoma MCF-7 and T-Lymphoblastoid Cells After Treatment with Herceptin and Herceptin/Lipoplex

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