Journal
BIOLOGICAL CHEMISTRY
Volume 390, Issue 1, Pages 11-18Publisher
WALTER DE GRUYTER GMBH
DOI: 10.1515/BC.2009.004
Keywords
Clostridium histolyticum; Clostridium tetani; enzymatic mechanism; FALGPA; inhibition; zinc metalloprotease
Categories
Funding
- Wissenschaftsfonds (FWF) [P20582]
- Austrian Science Fund (FWF) [P 20582] Funding Source: researchfish
Ask authors/readers for more resources
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 CoIH showing the highest overall activity and highest substrate affinity. Whereas the K. values of all three enzymes were within the same order of magnitude, the turnover rate k(cat) of ColG decreased 50- to 150-fold when compared to ColT and CoIH. 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.
Authors
I am an author on this paper
Click your name to claim this paper and add it to your profile.
Reviews
Recommended
No Data Available