Biochemical characterization of the catalytic domains of three different clostridial collagenases
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Ulrich Eckhard
Abstract
Clostridial collagenases are used for a broad spectrum of biotechnological applications and represent prime target candidates for both therapy and diagnosis of clostridial infections. In this study, we biochemically characterized the catalytic domains of three clostridial collagenases, collagenase G (ColG) and H (ColH) from Clostridium histolyticum, and collagenase T (ColT) from C. tetani. All protein samples showed activity against a synthetic peptidic substrate (furylacryloyl-Leu-Gly-Pro-Ala, FALGPA) with ColH showing the highest overall activity and highest substrate affinity. Whereas the Km values of all three enzymes were within the same order of magnitude, the turnover rate kcat of ColG decreased 50- to 150-fold when compared to ColT and ColH. It is noteworthy that the protein N-terminus significantly impacts their substrate affinity and substrate turnover as well as their inhibition profile with 1,10-phenanthroline. These findings were complemented with the discovery of a strictly conserved double-glycine motif, positioned 28 amino acids upstream of the HEXXH zinc binding site, which is critical for enzymatic activity. These observations have consequences with respect to the topology of the N-terminus relative to the active site as well as possible activation mechanisms.
©2009 by Walter de Gruyter Berlin New York
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Articles in the same Issue
- Editor's Note
- Editor's Note
- Protein Structure and Function
- Glyceryl ether monooxygenase resembles aromatic amino acid hydroxylases in metal ion and tetrahydrobiopterin dependence
- Biochemical characterization of the catalytic domains of three different clostridial collagenases
- Impact of detergents on the activity of acetylcholinesterase and on the effectiveness of its inhibitors
- The ADP-ribosylating thermozyme from Sulfolobus solfataricus is a DING protein
- Membranes, Lipids, Glycobiology
- Glycosphingolipids from bovine milk and milk fat globule membranes: a comparative study. Adhesion to enterotoxigenic Escherichia coli strains
- Mannose 6-phosphate receptor-dependent endocytosis of lysosomal enzymes is increased in sulfatide-storing kidney cells
- Cell Biology and Signaling
- Heavy metals induce phosphorylation of the Bcl-2 protein by Jun N-terminal kinase
- Fibroblast growth factor 2 (FGF-2) is a novel substrate for arginine methylation by PRMT5
- Differential functions of the Apoer2 intracellular domain in selenium uptake and cell signaling
- Active immunisation against gastric inhibitory polypeptide (GIP) improves blood glucose control in an animal model of obesity-diabetes
- Novel Techniques
- Applicability of superfolder YFP bimolecular fluorescence complementation in vitro
