Journal article
Analysis of heparan sulfate oligosaccharides by nano-electrospray ionization mass spectrometry
Glycobiology (Oxford), Vol.11(6), pp.505-513
06/01/2001
DOI: 10.1093/glycob/11.6.505
PMID: 11445555
Abstract
A highly sensitive method to identify and quantify heparan sulfate (HS) oligosaccharides by using nano-electrospray ionization mass spectrometry (nESI-MS) is described. The new approach allows us to detect approximately 50 nM of a chemically synthesized pentasaccharide with a structure of GlcNS6S-GlcA-GlcNS6S-IdoA2S-GlcNS6SOMe (3-OH pentasaccharide). Typically, solutions were infused for a total of 5 min, at an average flow rate of 30 nl/min, and the remaining sample was recovered from the nanovial. The spectra shown were obtained by summing scans for 1-3 min. Hence, our data indicated that as little as 3 x 10 -15 mole of the pentasaccharide was consumed to obtain a reasonable spectrum at the concentration as low as 50 nM. In addition, we found a linear relationship between the relative response of the molecular ion and the concentration of the analyzed 3-OH pentasaccharide, demonstrating that this approach can be used to determine the amount of HS oligosaccharides. To this end, a 3-O-sulfated pentasaccharide was prepared by incubating the 3-OH pentasaccharide with purified HS 3-O-sulfotransferase-1 and 3'-phosphoadenosine-5'-phospho[ 35S]sulfate. The resulting 3-O-sulfated pentasaccharide was purified and analyzed by nESI-MS. Based on the standard curve constructed with the 3-OH pentasaccharide, we calculated the concentration of the 3-O-sulfated pentasaccharide by the relative response. The result indicates that this value is very close to the value measured by [ 35S]sulfate radioactivity. In conclusion, nESI-MS provides both high sensitivity and the capacity to quantify HSs. This approach is likely to become a very important tool for structural analysis and sequencing of HS and heparin oligosaccharides.
Details
- Title: Subtitle
- Analysis of heparan sulfate oligosaccharides by nano-electrospray ionization mass spectrometry
- Creators
- R. Marshall Pope - University of North Carolina at Chapel HillChristina RaskaSuzanne ThorpJian Liu
- Resource Type
- Journal article
- Publication Details
- Glycobiology (Oxford), Vol.11(6), pp.505-513
- Publisher
- Oxford Publishing Limited(England)
- DOI
- 10.1093/glycob/11.6.505
- PMID
- 11445555
- ISSN
- 0959-6658
- eISSN
- 1460-2423
- Language
- English
- Date published
- 06/01/2001
- Academic Unit
- Medicine Administration
- Record Identifier
- 9984627202602771
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