Gas‐Liquid Chromatography and Mass Spectrometry in Methylation Analysis of Polysaccharides

Björndal, Håkan; Hellerqvist, Carl G.; Lindberg, Bengt; Svensson, Sigfrid

doi:10.1002/anie.197006101

Cited by 733 publications

(199 citation statements)

References 33 publications

Supporting

Mentioning

191

Contrasting

Order By: Relevance

“…Tentative identifications are listed in the figure legend. Qualitatively, the derivatives obtained are similar for all ages examined (5,8,12,14,16, and 20 DPA-data shown only for 8,12,16,and 20 DPA). Keeping in mind the limitations stated above, several conclusions can be drawn.…”

Section: Resultsmentioning

confidence: 63%

Changes in Biochemical Composition of the Cell Wall of the Cotton Fiber During Development

Meinert¹,

Delmer²

1977

Plant Physiol.

296

199

View full text Add to dashboard Cite

The composition of the cell wall of the cotton fiber (Gossypium hirsutum L. Acala SJ-1) has been studied from the early stages of elongation (5 days postaathesis) through the period of secondary wall formation, using cell wals derived both from fibers developing on the plant and from fibers obtained from excised, cultured ovules. The cell wall of the elongating cotton fiber was shown to be a dynamic structure.

show abstract

Section: Resultsmentioning

confidence: 63%

Changes in Biochemical Composition of the Cell Wall of the Cotton Fiber During Development

Meinert¹,

Delmer²

1977

Plant Physiol.

296

199

View full text Add to dashboard Cite

show abstract

“…The neutral and amino glycosyl compositions were determined by the alditol acetate method (1, I 1). Glycosyl-linkage compositions were determined by GC and GC-MS analysis of the partially methylated alditol acetate derivatives (9,28). Polysaccharide methylations were performed using modifications (11,28,30) of the procedure reported by Hakomori (18.…”

Section: Methodsmentioning

confidence: 99%

Structure of Plant Cell Walls

McNeil¹,

Darvill²,

Albersheim³

1980

Plant Physiol.

231

View full text Add to dashboard Cite

The purification and characterization of a pectic polymer, rhamnogalacturonan I, present in the primary cell walls of dicots is described. Rhamnogalacturonan I accounts for approximately 7% of the mass of the walls isolated from suspension-cultured sycamore cells. As purified, rhamnogalacturonan I has a molecular weight of approximately 200,000 and is composed primarily of L-rhamnosyl, D-galacturonosyl, L-arabinosyl, and Dgalactosyl residues. The backbone of rhamnogalacturonan I is thought to be composed predominantly of D-galacturonosyl and L-rhamnosyl residues in a ratio of approximately 2:1. About half of the L-rhamnosyl residues are 2-linked and are glycosidically attached to C4 of a D-galacturonosyl residue.The other half of the L-rhamnosyl residues are 2,4-inked and have a Dgalacturonosyl residue glycosidically attached at C2. Sidechains averaging 6 residues in length are attached to C4 of the L-rhamnosyl residues. There are many different sidechains, containing variously linked L-arabinosyl, and/or n-galactosyl residues.Pectic polysaccharides are defined as those containing 4-linkeda-D-galacturonosyl residues (25). Pectic polysaccharides are important components of the primary cell walls of dicots, accounting for approximately 30% of the walls (8,20,25,28 Enzyme Purification. Endo-a-1,4-polygalacturonase was purified from Colletotrichum lindemuthianum (15) and checked for purity as described (I 1).Enzymic Extraction of Pectic Polysaccharides. The pectic polysaccharides were extracted from isolated sycamore cell walls by C. lindemuthianum endopolygalacturonase by the procedures de-Gel Filtration Chromatography. Gel filtration chromatography was performed on a 1.4-x 40-cm agarose A-1.Sm column or on a 3-x 50-cm agarose A-Sm column. The columns were equilibrated and eluted with 50 mm Na-acetate (pH 5.2). The void and included volumes of the columns were determined with blue dextran (Sigma) and NaCl, respectively. In some cases (as noted in the text), the agarose A-1.5m column was equilibrated and eluted with 50 mm Na-acetate ( Analysis of Glycosyl and Glycosyl-linkage Compositions. The neutral and amino glycosyl compositions were determined by the alditol acetate method (1, I 1). Glycosyl-linkage compositions were determined by GC and GC-MS analysis of the partially methylated alditol acetate derivatives (9, 28). Polysaccharide methylations were performed using modifications (11,28,30)

show abstract

“…Foremost has been GC/MS of samples following methylation, hydrolysis, reduction, and acetylation. Analysis of these products (methylated alditol acetates) remains a major supporting component to MS/MS analysis, 2 although the combined data sets rarely provide adequate information for an exacting definition of structure. Thus, such proposed sequences are supplemented with intuition and biological insight, not physical measurement, exposing opportunities for misinterpretation.…”

mentioning

confidence: 99%

Carbohydrate Structural Isomers Analyzed by Sequential Mass Spectrometry

et al. 2007

View full text Add to dashboard Cite

Consistent with the goals of a comprehensive carbohydrate sequencing strategy, we extend earlier reports to include the characterization of structural (constitutional) isomers. Protocols were developed around ion trap instrumentation providing sequential mass spectrometry (MS n ) and supported with automation and related computational tools. These strategies have been built on the principle that for a single structure all product spectra upon sequential fragmentation are reproducible and each stage represents a rational spectrum of its precursor; i.e., all major fragments should be accounted for. Anomalous ions at any stage are clues indicating the presence of structural isomers. Gas-phase isolation and subsequent fragmentation of such ions provide an opportunity to specifically resolve selected structures for their detailed characterization. Importantly, some isomers were not detected following MS 2 and required multiple (MS n>2 ) stages for their characterization. Derivatization remains critical to position substructures in a glycan array since product ions carry fragmentation "scars" throughout the MS n tree. Equally as important are the pathway relationships between each stage and the greater yield of fragments with the smaller number of oscillators. Applications were directed to the structural isomers in ovalbumin and IgG, where, in the latter case, several previously unreported glycans were detected. Procedures were supported with bioinformatics tools for assimilating structure from the MS n data sets.A comprehensive carbohydrate sequence is particularly challenging when considering the constitutional and stereoisomers that are abundant in glycan arrays. Constitutional isomers, also referred to as structural isomers, are defined as an identical atomic composition arranged in a different structure. Stereoisomers are exemplified with those frequently encountered in the hexoses, mannose, galactose, and glucose, while the two G1 glycans found in IgG are representative of simple structural isomers. In this latter case, an identical monomer may be linked on either of two antennae providing a different topology. In another nomenclature clarification, the term isomer is often confused with isobar. which relates to a different composition of atoms occurring at a nominal (unit) mass resolution. 1 At the MS profile stage, isobars are not usually a significant problem in carbohydrate structural characterization, but structural isomers and stereoisomers are. During fragmentation, however, isobaric fragments may be formed, but identification will require higher mass resolving power and mass accuracy.

show abstract

Gas‐Liquid Chromatography and Mass Spectrometry in Methylation Analysis of Polysaccharides

Cited by 733 publications

References 33 publications

Changes in Biochemical Composition of the Cell Wall of the Cotton Fiber During Development

Changes in Biochemical Composition of the Cell Wall of the Cotton Fiber During Development

Structure of Plant Cell Walls

Carbohydrate Structural Isomers Analyzed by Sequential Mass Spectrometry

Contact Info

Product

Resources

About