Application

Chondroitinase ABC from Sigma has been used to digest O-linked chondroitin sulfate-like glycosaminoglycan (CSGAG). The enzyme has also been used to digest glycosaminoglycan chains in proteoglycans during the construction and expression of a recombinant versican gene in mammalian systems. Furthermore, it has been used to digest chondroitin sulfate in the proteoglycan complexes. The enzyme has also been used in the removal of glycan chains from aggrecan, which eliminates the axon growth-inhibitory effect of aggrecan.

Biochem/physiol Actions

Chondroitinase ABC catalyzes the eliminative degradation of polysaccharides containing (1-4)-β-D-hexosaminyl and (1-3)-β-D-glucuronosyl (or (1-3)-A-L-iduronosyl) linkages to disaccharides containing 4-deoxy-β-D-gluc-4-enuronosyl groups. It acts on chondroitin 4-sulfate, chondroitin 6-sulfate, and dermatan sulfate. However, it acts slowly on hyaluronate. The molecular weight is approximately 120 kDa with 2 non-identical subunits of molecular masses 86 kDa and 32 kDa. The pH optimum is 8.0 with chondroitin sulfate and 6.8 with hyaluronic acid. The optimal temperature is 37 °C. 1 mM Zn²⁺ acts as an inhibitor, while 0.05 M acetate is an activator of the enzyme.

Other Notes

View more information on enzymes for complex carbohydrate analysis at www.sigma-aldrich.com/enzymeexplorer

Packaging

2, 5, 10 units in glass bottle

Packages based on chondroitinase C activity.

Physical form

Lyophilized powder containing Tris buffer salts

Preparation Note

Reconstitute in a 0.01% bovine serum albumin aqueous (BSA) solution. Subsequent dilutions can be made into a buffer containing 50 mM Tris, pH 8.0, with 60 mM sodium acetate and 0.02% BSA. Solutions should be freshly prepared.

Unit Definition

One unit will liberate 1.0 μmole of a mixture of 2-acetamido-2-deoxy-3-O-(β-D-gluc-4-ene-pyranosyluronic acid)-4-O-sulfo-D-galactose and 1.0 μmole of 2-acetamido-2-deoxy-3-O-(β-D-gluc-4-ene-pyranosyluronic acid)-6-O-sulfo-D-galactose from chondroitin sulfate from shark cartilage, per min at pH 8.0 at 37 °C.