title: Insights into the mechanism of action of hyaluronate lyase: role of C-terminal domain and Ca2+ in the functional regulation of enzyme.
abstract: Hyaluronate lyases (HLs) cleave hyaluronan and certain other chondroitin/chondroitin sulfates. Although native HL from Streptococcus agalactiae is composed of four domains, it finally stabilizes after autocatalytic conversion as a 92-kDa enzyme composed of the N-terminal spacer, middle alpha-, and C-terminal domains. These three domains are independent folding/unfolding units of the enzyme. Comparative structural and functional studies using the enzyme and its various fragments/domains suggest a relatively insignificant role of the N-terminal spacer domain in the 92-kDa enzyme. Functional studies demonstrate that the alpha-domain is the catalytic domain. However, independently it has a maximum of only about 10% of the activity of the 92-kDa enzyme, whereas its complex with the C-terminal domain in vitro shows a significant enhancement (about 6-fold) in the activity. It has been previously proposed that the C-terminal domain modulates the enzymatic activity of HLs. In addition, one of the possible roles for calcium ions was suggested to induce conformational changes in the enzyme loops, making HL more suitable for catalysis. However, we observed that calcium ions do not interact with the enzyme, and its role actually is in modulating the hyaluronan conformation and not in the functional regulation of enzyme.
authors.affiliation: Central Drug Research Institute, Lucknow 226 001, India.
authors.forename: Md Sohail
authors.initials: MS
authors.lastname: Akhtar
authors.forename: Manju Yasoda
authors.initials: MY
authors.lastname: Krishnan
authors.forename: Vinod
authors.initials: V
authors.lastname: Bhakuni
mesh_terms: D002118:Calcium / Q000494:pharmacology*; D007700:Kinetics; D011133:Polysaccharide-Lyases / Q000737:chemistry* / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D013292:Streptococcus agalactiae / Q000201:enzymology*
chemical_list: D011133:Polysaccharide-Lyases; C100002:hyaluronate lyase; D002118:Calcium