Hyaluronidases Have Strong Hydrolytic Activity toward Chondroitin 4-Sulfate Comparable to that for Hyaluronan

Abstract

Chondroitin sulfate (CS) chains are involved in the regulation of various biological processes. However, the mechanism underlying the catabolism of CS is not well understood. Hyaluronan (HA)-degrading enzymes, the hyaluronidases, are assumed to act at the initial stage of the degradation process, because HA is similar in structure to nonsulfated CS, chondroitin (Chn). Although human hyaluronidase-1 (HYAL1) and testicular hyaluronidase (SPAM1) can degrade not only HA but also CS, they are assumed to digest CS to only a limited extent. In this study, the hydrolytic activities of HYAL1 and SPAM1 toward CS-A, CS-C, Chn, and HA were compared. HYAL1 depolymerized CS-A and HA to a similar extent. SPAM1 degraded CS-A, Chn, and HA to a similar extent. CS is widely distributed from very primitive organisms to humans, whereas HA has been reported to be present only in vertebrates with the single exception of a mollusk. Therefore, a genuine substrate of hyaluronidases appears to be CS as well as HA.

Inhibitory effect of chondroitin sulfate oligosaccharides on bovine testicular hyaluronidase

“…Testicular hyaluronidase has been known to show a preference towards HA over Ch4S as a hydrolysis substrate and that was consistent at pH 5, 37 • C (typical condition), whereas it showed a preference towards Ch4S over HA in selective conditions such as at pH 6, 45 • C (Knudson, Gundlach, Schmid, & Conrad, 1984). Recently, it was demonstrated that Ch4S is a better substrate than HA for testicular hyaluronidase (recombinant SPAM1) at pH 4 or lower pH, whereas HA is a better substrate than Ch4S above pH 4 (Honda, Kaneiwa, Mizumoto, Sugahara, & Yamada, 2012). In either condition in both reports Ch6S was a poorer substrate than HA and Ch4S.…”

“…In either condition in both reports Ch6S was a poorer substrate than HA and Ch4S. Their kinetic data suggested that testicular hyaluronidase has higher affinity for Ch4S and HA than for Ch6S (Honda et al, 2012). These findings may explain why that when both the chain length and sufation degree are similar Ch4S oligosaccharides are better inhibitors than Ch6S oligosaccharides for BTH from the aspect of the affinity with the enzyme.…”

Following the enzymatic digestion of chondroitin sulfate by a simple GPC analysis

“…Hyaluronidase (HASE) and chondroitinase (CASE) are two enzymes that are active upon CS in vivo [20][21][22]. We used both enzymes in our studies aiming validation of the proposed method for different substrates and enzymes.…”

Hyaluronidase-Sensitive Nanoparticle Templates for Triggered Release of HIV/AIDS Microbicide In Vitro

“…The optimum pH for maximum hydrolytic activity of HAase is 4 to 5 (37,38). However, it has been reported that HAase exert its activity at weakly acidic (pH 5 to 6) (38) and alkaline conditions (pH 7 to 8) (37,39). The bovine testicular HAase was used as a model enzyme, since it is commercially available and, like human HAase, it catalyzes the hydrolysis of the β(1-4) glycosidic bonds in HA (40).…”