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. 2009 May 1;284(18):11806-14.
doi: 10.1074/jbc.M809637200. Epub 2009 Feb 25.

Organ-specific heparan sulfate structural phenotypes

Xiaofeng Shi  1 , Joseph Zaia
Affiliations

Organ-specific heparan sulfate structural phenotypes

Xiaofeng Shi et al. J Biol Chem. .

Abstract

The functions of heparan sulfate (HS) depend on the expression of structural domains that interact with protein partners. Glycosaminoglycans (GAGs) exhibit a high degree of polydispersity in their composition, chain length, sulfation, acetylation, and epimerization patterns. It is essential for the understanding of GAG biochemistry to produce detailed structural information as a function of spatial and temporal factors in biological systems. Toward this end, we developed a set of procedures to extract GAGs from various rat organ tissues and examined and compared HS expression levels using liquid chromatography/mass spectrometry. Here we demonstrate detailed variations in HS GAG chains as a function of organ location. These studies shed new light on the structural variation of GAG chains with respect to average length, disaccharide composition, and expression of low abundance structural epitopes, including unsubstituted amino groups and lyase-resistant oligosaccharides. The data show the presence of a disaccharide with an unsubstituted amino group that is endogenous and widely expressed in mammalian organ tissues.

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Figures

FIGURE 1.
FIGURE 1.
Extracted HS dp2 ion chromatogram from rat brain tissues analyzed by SEC LC/MS. Top, 4,5-unsaturated dp2; bottom, saturated dp2. 1, ΔHexA-GlcNAc; 2, ΔHexA-GlcNS; 3, ΔHexA(2S)-GlcNAc (6S); 4, ΔHexA(2S)-GlcNS (6S); 5, ΔHexA2S-GlcNAc6S; 6, ΔHexA2S-NS6S; 1′, HexA-GlcNAc; 2′, HexA-GlcNS; 3′, HexA(2S)-GlcNAc (6S); 4′, HexA(2S)-GlcNS (6S); 5′, HexA2S-GlcNAc6S; 6′, HexA2S-GlcNS6S.
FIGURE 2.
FIGURE 2.
HS disaccharide profiles in rat tissues analyzed by SEC LC/MS. The percentage of disaccharides of total digest abundance was as follows: heart, 95.7%; kidney, 92.5%; lung, 96.2%; spleen, 91.4%; brain, 84.9%; liver, 95.3%; muscle, 96.1%.
FIGURE 3.
FIGURE 3.
The average number of sulfations per 100 dp2s among rat tissues.
FIGURE 4.
FIGURE 4.
Average chain lengths of HS in rat tissues calculated by unsaturated and saturated disaccharides.
FIGURE 5.
FIGURE 5.
The probabilities of HS disaccharides to be at the nonreducing end position among rat tissues.
FIGURE 6.
FIGURE 6.
A plot of the logarithm of the ratio of the probabilities of HS disaccharides at the nonreducing end and in the entire chain.
FIGURE 7.
FIGURE 7.
The identification of unsubstituted ΔHexA (2S)-GlcNH2 (6S) from tissue samples.
FIGURE 8.
FIGURE 8.
The distribution of ΔHexA-GlcNH26S from different rat tissues. ND, abundance not intense enough to be determined.
FIGURE 9.
FIGURE 9.
Lyase-resistant oligosaccharide distribution in different rat tissues.
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