Chondroitin sulfate (CS) is widely distributed as the glycosaminoglycan side chains of proteoglycans in extracellular matrices and at the cell surface. CS chains have a linear structure composed of repeating disaccharide units comprising d-glucuronic acid and N-acetyl-d-galactosamine, which are sulfated at different positions in various combinations. This structural diversity is responsible for the multiple biological functions of CS.
The cellular degradation of CS occurs predominantly in lysosomes. Following the fragmentation of polysaccharides by endo-type hydrolases, the oligosaccharide products are degraded sequentially from the nonreducing end by exo-type glycosidases and sulfatases, although no endoglycosidases specific to CS have been reported. In this study, human hyaluronidase-4 was demonstrated to be a CS-specific endo-beta-N-acetylgalactosaminidase. The specificity of a purified recombinant form of the enzyme was investigated using various CS isoforms, and the structure of the best cleavage site was characterized.
This enzyme will be a useful tool for investigating CS-specific functions in tissues and cells without degrading hyaluronan. It may also be applicable to the treatment of acute spinal cord injuries instead of the bacterial CS lyase used in recent clinical trials.
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
High performance liquid chromatography
This is a preview of subscription content, log in to check access.
This work was supported in part by a Grant-in-aid for Scientific Research (C) (21590057) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan (MEXT), and the Mizutani Foundation for Glycoscience, Tokyo, Japan. I thank Professor Kazuyuki Sugahara for his helpful suggestions as well as Tomoyuki Kneiwa, Anzu Miyazaki, and Shuji Mizumoto for their many contributions.
Csóka AB, Scherer SW, Stern R (1999) Expression analysis of six paralogous human hyaluronidase genes clustered on chromosomes 3p21 and 7q31. Genomics 60:356–361PubMedCrossRefGoogle Scholar
Csoka AB, Frost GI, Stern R (2001) The six hyaluronidase-like genes in the human and mouse genomes. Matrix Biol 20:499–508PubMedCrossRefGoogle Scholar
Duterme C, Mertens-Strijthagen J, Tammi MI, Flamion B (2009) Two novel functions of hyaluronidase-2 (Hyal2): formation of the glycocalyx and control of CD44-ERM interactions. J Biol Chem 284:33495–33508PubMedCrossRefGoogle Scholar
Esko JD, Selleck SB (2002) Order out of chaos: assembly of ligand binding sites in heparan sulfate. Annu Rev Biochem 71:435–471PubMedCrossRefGoogle Scholar
Harada H, Takahashi M (2007) CD44-dependent intracellular and extracellular catabolism of hyaluronic acid by hyaluronidase-1 and -2. J Biol Chem 282:5597–5607PubMedCrossRefGoogle Scholar
Jedrzejas MJ, Stern R (2005) Structures of vertebrate hyaluronidases and their unique enzymatic mechanism of hydrolysis. Proteins 61:227–238PubMedCrossRefGoogle Scholar
Kaneiwa T, Yamada S, Mizumoto S, Montaño AM, Mitani S, Sugahara K (2008) Identification of a novel chondroitin hydrolase in Caenorhabditis elegans. J Biol Chem 283:14971–14979PubMedCrossRefGoogle Scholar
Kaneiwa T, Mizumoto S, Sugahara K, Yamada S (2010) Identification of human hyaluronidase-4 as a novel chondroitin sulfate hydrolase that preferentially cleaves the galactosaminidic linkage in the trisulfated tetrasaccharide sequence. Glycobiology 20:300–309PubMedCrossRefGoogle Scholar
Lepperdinger G, Strobl B, Kreil G (1998) HYAL2, a human gene expressed in many cells, encodes a lysosomal hyaluronidase with a novel type of specificity. J Biol Chem 273:22466–22470PubMedCrossRefGoogle Scholar
Prabhakar V, Sasisekharan R (2006) The biosynthesis and catabolism of galactosaminoglycans. Adv Pharmacol 53:69–115PubMedCrossRefGoogle Scholar
Rai SK, Duh FM, Vigdorovich V, Danilkovitch-Miagkova A, Lerman MI, Miller AD (2001) Candidate tumor suppressor HYAL2 is a glycosylphosphatidylinositol (GPI)-anchored cell-surface receptor for jaagsiekte sheep retrovirus, the envelope protein of which mediates oncogenic transformation. Proc Natl Acad Sci USA 98:4443–4448PubMedCrossRefGoogle Scholar
Rauch U, Kappler L (2006) Chondroitin/dermatan sulfates in the central nervous system: their structures and functions in health and disease. Adv Pharmacol 53:337–356PubMedCrossRefGoogle Scholar
Rodén L (1980) Structure and metabolism of connective tissue proteoglycans. In: Lennarz WJ (ed) The biochemistry of glycoproteins and poteoglycans. Plenum, New York, pp 267–371CrossRefGoogle Scholar
Sugahara K, Yamada S (2000) Structure and function of oversulfated chondroitin sulfate variants: unique sulfation patterns and neuroregulatory activities. Trends Glycosci Glycotechnol 12:321–349CrossRefGoogle Scholar
Sugahara K, Mikami T, Uyama T, Mizuguchi S, Nomura K, Kitagawa H (2003) Recent advances in the structural biology of chondroitin sulfate and dermatan sulfate. Curr Opin Struct Biol 13:612–620PubMedCrossRefGoogle Scholar
Uyama T, Kitagawa H, Sugahara K (2007) Biosynthesis of glycosaminoglycans and proteoglycans. In: Kamerling JP (ed) Comprehensive glycoscience, vol 3. Elsevier, Amsterdam, pp 79–104CrossRefGoogle Scholar
Yamada S, Sugahara K (2008) Potential therapeutic application of chondroitin sulfate/dermatan sulfate. Curr Drug Discov Technol 5:289–301PubMedCrossRefGoogle Scholar
Yamada S, Van Die I, Van den Eijnden DH, Yokota A, Kitagawa H, Sugahara K (1999) Demonstration of glycosaminoglycans in Caenorhabditis elegans. FEBS Lett 459:327–331PubMedCrossRefGoogle Scholar
Yamada S, Mizumoto S, Sugahara K (2009) Chondroitin hydrolase in Caenorhabditis elegans. Trends Glycosci Glycotechnol 21:149–162CrossRefGoogle Scholar
Yamada S, Sugahara K, Özbek S (2011) Evolution of glycosaminoglycans: comparative biochemical study. Commun Integr Biol 4(2):150–8PubMedCrossRefGoogle Scholar