Electron detachment dissociation of synthetic heparan sulfate glycosaminoglycan tetrasaccharides varying in degree of sulfation and hexuronic acid stereochemistry

Leach, Franklin E.; Arungundram, Sailaja; Al-Mafraji, Kanar; Venot, André; Boons, Geert‐Jan; Amster, I. Jonathan

doi:10.1016/j.ijms.2012.07.002

Cited by 27 publications

(37 citation statements)

References 45 publications

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“…Close examination of the EDD spectra for the GlcA containing standards 1a and 1b, Figure 4a, reveals results consistent with previous EDD reports on modestly sulfated HS tetrasaccharides [35, 40]. Diagnostic ion B 3 ′-CO 2 indicative of glucuronic acid in less sulfated HS GAGs (0–0.25 sulfates per disaccharide) reported for naturally extracted and synthetically produced HS tetrasaccharides [18, 21] are observed for only samples 1a (IdoA2S-GlcNAc-GlcA-GlcNAc-(CH 2 ) 5 NH 2) and 1b (IdoA-GlcNAc6S-GlcA-GlcNAc-(CH 2 ) 5 NH 2) Figure 4a, occurring at m/z 589.0972. An expanded mass spectra region for the stereospecific ion B 3 ′-CO 2 is shown in Figure 4a.…”

Section: Resultssupporting

confidence: 87%

“…Previous EDD and CID reports have shown the presence of sodium counter ions can provide a means to control stereochemical dependent fragmentation [21, 36]. We found that to be very useful for the tri and tetra-sulfated HS standards.…”

Section: Resultsmentioning

confidence: 80%

“…Negative electrospray ionization mass spectrometry offers an excellent platform for GAG analysis offering high sensitivity, throughput, and accuracy [19, 20]. However, tandem mass spectrometry of GAGs especially heparan sulfate is often challenging due to variations in oligomer length, hexuronic acid stereochemistry, sulfation heterogeneity[6, 21]. ESI-MS is able to determine the length, degree of sulfation of GAGs, and other features affecting the elemental composition.…”

Section: Introductionmentioning

confidence: 99%

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Single Stage Tandem Mass Spectrometry Assignment of the C-5 Uronic Acid Stereochemistry in Heparan Sulfate Tetrasaccharides using Electron Detachment Dissociation

Agyekum

Zong

Boons

et al. 2017

J. Am. Soc. Mass Spectrom.

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The analysis of heparan sulfate (HS) glycosaminoglycans presents many challenges, due to the high degree of structural heterogeneity arising from their non-template biosynthesis. Complete structural elucidation of glycosaminoglycans necessitates the unambiguous assignments of sulfo modifications and the C-5 uronic acid stereochemistry. Efforts to develop tandem mass spectrometric-based methods for the structural analysis of glycosaminoglycans have focused on the assignment of sulfo positions. The present work focuses on the assignment of the C-5 stereochemistry of the uronic acid that lies closest to the reducing end. Prior work with electron-based tandem mass spectrometry methods, specifically, electron detachment dissociation (EDD), have shown great promise in providing stereo-specific product ions, such as the B3′–CO2, which has been found to distinguish glucuronic acid (GlcA) from iduronic acid (IdoA) in some HS tetrasaccharides. The previously observed diagnostic ion are generally not observed with 2-O-sulfo uronic acids or for more highly sulfated heparan sulfate tetrasaccharides. A recent study using electron detachment dissociation and principal component analysis revealed a series of ions that correlate with GlcA versus IdoA for a set of 2-O-sulfo HS tetrasaccharide standards. This present work comprehensively investigates the efficacy of these ions for assigning the C-5 stereochemistry of the reducing end uronic acid in thirty-three HS tetrasaccharides. A diagnostic ratio can be computed from the sum of the ions that correlate to GlcA to those that correlate to IdoA.

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Section: Resultssupporting

confidence: 87%

Section: Resultsmentioning

confidence: 80%

Section: Introductionmentioning

confidence: 99%

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Single Stage Tandem Mass Spectrometry Assignment of the C-5 Uronic Acid Stereochemistry in Heparan Sulfate Tetrasaccharides using Electron Detachment Dissociation

Agyekum

Zong

Boons

et al. 2017

J. Am. Soc. Mass Spectrom.

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“…EDD has been applied to most sulfated GAG glycoform classes including chondroitin/dermatan sulfates, heparan sulfate, and heparin ranging from tetramers to decamers [11, 25, 27, 39, 48]. EDD consists of two simultaneous/coupled pathways that result in fragment ion production.…”

Section: Resultsmentioning

confidence: 99%

Negative Electron Transfer Dissociation Sequencing of Increasingly Sulfated Glycosaminoglycan Oligosaccharides on an Orbitrap Mass Spectrometer

Leach

Riley

Westphall

et al. 2017

J. Am. Soc. Mass Spectrom.

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The structural characterization of sulfated glycosaminoglycan (GAG) carbohydrates remains an important target for analytical chemists due to challenges introduced by the natural complexity of these mixtures and the defined need for molecular-level details to elucidate biological structure-function relationships. Tandem mass spectrometry has proven the most powerful technique for this purpose. Previously, electron detachment dissociation (EDD), in comparison to other methods of ion activation, has been shown to provide the largest number of useful cleavages for de novo sequencing of GAG oligosaccharides, but such experiments are restricted to Fourier transform ion cyclotron resonance mass spectrometers (FTICR-MS). Negative electron transfer dissociation (NETD) provides similar fragmentation results, and can be achieved on any mass spectrometry platform that is designed to accommodate ion-ion reactions. Here, we examine for the first time the effectiveness of NETD-Orbitrap mass spectrometry for the structural analysis of GAG oligosaccharides. Compounds ranging in size from tetrasaccharides to decasaccharide were dissociated by NETD, producing both glycosidic and cross-ring cleavages that enabled the location of sulfate modifications. The highly-sulfated, heparin-like synthetic GAG, Arixtra™, was also successfully sequenced by NETD. In comparison to other efforts to sequence GAG chains without fully ionized sulfate constituents, the occurrence of sulfate loss peaks is minimized by judicious precursor ion selection. The results compare quite favorably to prior results with electron detachment dissociation (EDD). Significantly, the duty cycle of the NETD experiment is sufficiently short to make it an effective tool for on-line separations, presenting a straightforward path for selective, high-throughput analysis of GAG mixtures.

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“…Despite their advantages, these fragmentation techniques were only reported to work well for HS samples with relatively low degrees of sulfation. 28,29 As the number of potential deprotonation sites per residue increases in highly sulfated HS, the precursor ion charge density also increases, leading to reduced anion/electron collision cross-section due to increased columbic repulsion. Consequently, electron detachment becomes less efficient and the tandem mass spectra are less useful for sequencing.…”

Section: Introductionmentioning

confidence: 99%

De Novo Sequencing of Heparan Sulfate Oligosaccharides by Electron-Activated Dissociation

Huang

Mao

et al. 2013

Anal. Chem.

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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) time-scale, 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.

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Electron detachment dissociation of synthetic heparan sulfate glycosaminoglycan tetrasaccharides varying in degree of sulfation and hexuronic acid stereochemistry

Cited by 27 publications

References 45 publications

Single Stage Tandem Mass Spectrometry Assignment of the C-5 Uronic Acid Stereochemistry in Heparan Sulfate Tetrasaccharides using Electron Detachment Dissociation

Single Stage Tandem Mass Spectrometry Assignment of the C-5 Uronic Acid Stereochemistry in Heparan Sulfate Tetrasaccharides using Electron Detachment Dissociation

Negative Electron Transfer Dissociation Sequencing of Increasingly Sulfated Glycosaminoglycan Oligosaccharides on an Orbitrap Mass Spectrometer

De Novo Sequencing of Heparan Sulfate Oligosaccharides by Electron-Activated Dissociation

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