Forskning ved Københavns Universitet - Københavns Universitet

Forside

De novo sequencing of heparan sulfate oligosaccharides by electron-activated dissociation

Publikation: Bidrag til tidsskriftTidsskriftartikelForskningfagfællebedømt

  • Yu Huang
  • Xiang Yu
  • Yang Mao
  • Catherine E Costello
  • Joseph Zaia
  • Cheng Lin

Structural characterization of highly sulfated glycosaminoglycans (GAGs) by collisionally activated dissociation (CAD) is challenging because of the extensive sulfate losses mediated by free protons. While removal of the free protons may be achieved through the use of derivatization, metal cation adducts, and/or electrospray supercharging reagents, these steps add complexity to the experimental workflow. It is therefore desirable to develop an analytical approach for GAG sequencing that does not require derivatization or addition of reagents to the electrospray solution. Electron detachment dissociation (EDD) can produce extensive and informative fragmentation for GAGs without the need to remove free protons from the precursor ions. However, EDD is an inefficient process, often requiring consumption of large sample quantities (typically several micrograms), particularly for highly sulfated GAG ions. Here, we report that with improved instrumentation, optimization of the ionization and ion transfer parameters, and enhanced EDD efficiency, it is possible to generate highly informative EDD spectra of highly sulfated GAGs on the liquid chromatography (LC) timescale, with consumption of only a few nanograms of sample. We further show that negative electron transfer dissociation (NETD) is an even more effective fragmentation technique for GAG sequencing, producing fewer sulfate losses while consuming smaller amount of samples. Finally, a simple algorithm was developed for de novo HS sequencing based on their high-resolution tandem mass spectra. These results demonstrate the potential of EDD and NETD as sensitive analytical tools for detailed, high-throughput, de novo structural analyses of highly sulfated GAGs.

OriginalsprogEngelsk
TidsskriftAnalytical Chemistry
Vol/bind85
Udgave nummer24
Sider (fra-til)11979-86
Antal sider8
ISSN0003-2700
DOI
StatusUdgivet - 17 dec. 2013
Eksternt udgivetJa

ID: 161581258