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Isolation and characterization of heparan sulfate from various murine tissues

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Abstract

Heparan sulfate (HS), is a proteoglycan (PG) found both in the extracellular matrix and on cell surface. It may represent one of the most biologically important glycoconjugates, playing an essential role in a variety of different events at molecular level. The publication of the mouse genome, and the intensive investigations aimed at understanding the proteome it encodes, has motivated us to initiate studies in mouse glycomics focused on HS. The current study is aimed at determining the quantitative and qualitative organ distribution of HS in mice. HS from brain, eyes, heart, lung, liver, kidney, spleen, intestine and skin was purified from 6–8 week old male and female mice. The recovered yield of HS from these organs is compared with the recovered whole body yield of HS. Structural characterization of the resulting HS relied on disaccharide analysis and 1H-NMR spectroscopy. Different organs revealed a characteristic HS structure. These data begin to provide a structural understanding of the role of HS in cell-cell interactions, cell signaling and sub-cellular protein trafficking as well as a fundamental understanding of certain aspects of protein-carbohydrate interactions.

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Abbreviations

GAGs:

glycosaminoglycans

PG:

proteoglycans

HS:

heparan sulfate

SPR:

surface plasmon resonance

MWCO:

molecular weight cut-off

ΔUA-GlcNAc:

2-acetamido-2-deoxy-4-O-(4-deoxy-α-L-threo-hex-enepyranosyluronic acid)-D-glucose

ΔUA-GlcNS:

2-deoxy-2-sulfamido-4-O-(4-deoxy-α-L-threo-hex-enepyranosyluronic acid)-D-glucose

ΔUA-GlcNAc6S:

2-acetamido-2-deoxy-4-O-(4-deoxy-α-L-threo-hex-enepyranosyluronic acid)-6-O-sulfo- D-glucose

ΔUA2S-GlcNAc:

2-acetamido-2-deoxy-4-O-(4-deoxy-2-O-sulfo-α-L-threo-hex-enepyranosyluronic acid)-D-glucose

ΔUA-GlcNS6S:

2-deoxy-2-sulfamido-4-O-(4-deoxy-2-O-sulfo-α-L-threo-hex-enepyranosyluronic acid)-6-O-sulfo- D-glucose

ΔUA2S-GlcNS:

2-deoxy-2-sulfamido-4-O-(4-deoxy-2-O-sulfo-α-L-threo-hex-enepyranosyluronic acid)-D-glucose

ΔUA2S-GlcNAc6S:

2-acetamido-2-deoxy-4-O-(4-deoxy-2-O-sulfo-α-L-threo-hex-enepyranosyluronic acid)-6-O-sulfo-D-glucose

ΔUA2S-GlcNS6S:

2-deoxy-2-sulfamido-4-O-(4-deoxy-2-O-sulfo-α-L-threo-hex-enepyranosyluronic acid)-6-O-sulfo- D-glucose

CPC:

cetylpyridinium chloride

COSY:

correlation spectroscopy

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Correspondence to Robert J. Linhardt.

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Warda, M., Toida, T., Zhang, F. et al. Isolation and characterization of heparan sulfate from various murine tissues. Glycoconj J 23, 555–563 (2006). https://doi.org/10.1007/s10719-006-7668-1

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  • DOI: https://doi.org/10.1007/s10719-006-7668-1

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