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The effect of substitution of the N-acetyl groups of N-acetylgalactosamine residues in chondroitin sulfate on its degradation by chondroitinase ABC

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Abstract

Chondroitinase ABC is a lyase that degrades chondroitin sulfate, dermatan sulfate and hyaluronic acid into disaccharides. The purpose of this study was to determine the ability of chondroitinase ABC to degrade chondroitin sulfate in which the N-acetyl groups are substituted with different acyl groups. The bovine tracheal chondroitin sulfate A (bCSA) was N-deacetylated by hydrazinolysis, and the free amino groups derivatized into N-formyl, N-propionyl, N-butyryl, N-hexanoyl or N-benzoyl amides. Treatment of the N-acyl or N-benzoyl derivatives of bCSA with chondroitinase ABC and analysis of the products showed that the N-formyl, N-hexanoyl and N-benzoyl derivatives are completely resistant to the enzyme. In contrast, the N-propionyl or N-butyryl derivatives were degraded into disaccharides with slower kinetics compared to that of unmodified bCSA. The rate of degradation of bCSA derivatives by the enzyme was found to be in the order of N-acetyl>N-propionyl>>N-butyryl bCSA. These results have important implications for understanding the interaction of N-acetyl groups of glycosaminoglycans with chondroitinase ABC.

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Abbreviations

CS:

chondroitin sulfate

C4S:

chondroitin 4-sulfate

C6S:

chondroitin 6-sulfate

DS:

dermatan sulfate

HA:

hyaluronic acid

GAG:

glycosaminoglycan

CSPG:

chondroitin sulfate proteoglycan

DSPG:

dermatan sulfate proteoglycan

bCSA:

bovine tracheal chondroitin sulfate A

GlcA:

glucuronic acid

IdA:

iduronic acid

GalNAc:

N-acetylgalactosamine

GlcNAc:

N-acetylglucosamine

Gal:

galactose

Xyl:

xylose

Glc:

glucose

IRBCs:

infected red blood cells

BD:

blue dextran

EDAC:

N-ethyl-N′-(3-dimethylaminopropyl)carbodiimide

MES:

2-(N-morpholino)ethanesulfonic acid

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Acknowledgements

This work was supported by grant AI45086 from the National Institute of Allergy and Infectious Diseases, National Institutes of Health.

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Author notes

  1. SubbaRao V. Madhunapantula & Rajeshwara N. Achur

    Present address: Department of Pharmacology, Pennsylvania State University College of Medicine, Hershey, PA, 17033, USA

  2. Veer P. Bhavanandan

    Present address: Center for Glycosciences and Technology, The Biodesign Institute, Arizona State University, Tempe, AZ, 85287, USA

Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, Pennsylvania State University College of Medicine, 500 University Drive, Hershey, PA, 17033, USA

    SubbaRao V. Madhunapantula, Rajeshwara N. Achur, Veer P. Bhavanandan & D. Channe Gowda

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  1. SubbaRao V. Madhunapantula
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Correspondence to D. Channe Gowda.

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Madhunapantula, S.V., Achur, R.N., Bhavanandan, V.P. et al. The effect of substitution of the N-acetyl groups of N-acetylgalactosamine residues in chondroitin sulfate on its degradation by chondroitinase ABC. Glycoconj J 24, 465–473 (2007). https://doi.org/10.1007/s10719-007-9039-y

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