Dataset Information

Discrimination of large maltooligosaccharides from isobaric dextran and pullulan using ion mobility mass spectrometry.

ABSTRACT:

Rationale

Ion mobility mass spectrometry (IMMS) has previously been shown to resolve small isobaric oligosaccharides, but larger alpha-oligoglucans are also abundant in biology and are of industrial importance. If conformational differences between such isomers are retained in the gas phase, IMMS could be used to address questions in biology and industry.

Methods

Negative mode electrospray ionization (ESI) travelling-wave IMMS was used to resolve large isobaric ?-glucan ions on the basis of their different gas-phase conformations. ?,?-Dicarboxy-terminated polystyrene was used to calibrate the instrument allowing the collision cross-sections (CCSs) of ions to be determined.

Results

?-1,4-Linked maltooligosaccharides with a degree of polymerisation of up to 35 could be discriminated from ?-1,6-linked dextran and ?-1,4/1,6-linked pullulan using IMMS. Fragmentation spectra of ions separated by IMMS could also distinguish isomers. Two conformational isomers of maltohexaose were resolvable by IMMS, likely reflecting extended and V6 helical conformations. IMMS was also able to identify a product within a mixture of maltooligosaccharides treated with the potential anti-tuberculosis drug target Mycobacterium tuberculosis GlgB branching enzyme.

Conclusions

Biological samples of complex isobaric oligosaccharides can be analysed using IMMS in the negative mode providing facile analyses and high sensitivity without the need for either derivatisation or chromatographic separation.

SUBMITTER: Rashid AM

PROVIDER: S-EPMC4285287 | biostudies-literature | 2014 Jan

REPOSITORIES: biostudies-literature

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Publications

Discrimination of large maltooligosaccharides from isobaric dextran and pullulan using ion mobility mass spectrometry.

Rashid Abdul M AM Saalbach Gerhard G Bornemann Stephen S

Rapid communications in mass spectrometry : RCM 20140101 2

<h4>Rationale</h4>Ion mobility mass spectrometry (IMMS) has previously been shown to resolve small isobaric oligosaccharides, but larger alpha-oligoglucans are also abundant in biology and are of industrial importance. If conformational differences between such isomers are retained in the gas phase, IMMS could be used to address questions in biology and industry.<h4>Methods</h4>Negative mode electrospray ionization (ESI) travelling-wave IMMS was used to resolve large isobaric α-glucan ions on th ...[more]

PMID: 24338967

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Project description:In this report, we explored the benefits of cyclic ion mobility (cIM) mass spectrometry in the analysis of isomeric post-transcriptional modifications of RNA. Standard methyl-cytidine samples were initially utilized to test the ability to correctly distinguish different structures sharing the same elemental composition and thus molecular mass. Analyzed individually, the analytes displayed characteristic arrival times (tD ) determined by the different positions of the modifying methyl groups onto the common cytidine scaffold. Analyzed in mixture, the widths of the respective signals resulted in significant overlap that initially prevented their resolution on the tD scale. The separation of the four isomers was achieved by increasing the number of passes through the cIM device, which enabled to fully differentiate the characteristic ion mobility behaviors associated with very subtle structural variations. The placement of the cIM device between the mass-selective quadrupole and the time-of-flight analyzer allowed us to perform gas-phase activation of each of these ion populations, which had been first isolated according to a common mass-to-charge ratio and then separated on the basis of different ion mobility behaviors. The observed fragmentation patterns confirmed the structures of the various isomers thus substantiating the benefits of complementing unique tD information with specific fragmentation data to reach more stringent analyte identification. These capabilities were further tested by analyzing natural mono-nucleotide mixtures obtained by exonuclease digestion of total RNA extracts. In particular, the combination of cIM separation and post-mobility dissociation allowed us to establish the composition of methyl-cytidine and methyl-adenine components present in the entire transcriptome of HeLa cells. For this reason, we expect that this technique will benefit not only epitranscriptomic studies requiring the determination of identity and expression levels of RNA modifications, but also metabolomics investigations involving the analysis of natural extracts that may possibly contain subsets of isomeric/isobaric species.

Dataset Information

Discrimination of large maltooligosaccharides from isobaric dextran and pullulan using ion mobility mass spectrometry.

Rationale

Methods

Results

Conclusions

Publications

Discrimination of large maltooligosaccharides from isobaric dextran and pullulan using ion mobility mass spectrometry.

Similar Datasets

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