Document Detail

Biochemical characterization of the catalytic domains of three different Clostridial collagenases.
MedLine Citation:
PMID:  18937627     Owner:  NLM     Status:  MEDLINE    
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 K(m) 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 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.
Ulrich Eckhard; Esther Schönauer; Paulina Ducka; Peter Briza; Dorota Nüss; Hans Brandstetter
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biological chemistry     Volume:  390     ISSN:  1431-6730     ISO Abbreviation:  Biol. Chem.     Publication Date:  2009 Jan 
Date Detail:
Created Date:  2008-12-17     Completed Date:  2009-03-12     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9700112     Medline TA:  Biol Chem     Country:  Germany    
Other Details:
Languages:  eng     Pagination:  11-8     Citation Subset:  IM    
Division of Structural Biology, Department of Molecular Biology, University of Salzburg, Billrothstrasse 11, A-5020 Salzburg, Austria.
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MeSH Terms
Amino Acid Motifs
Amino Acid Sequence
Catalytic Domain*
Clostridium histolyticum / enzymology*
Clostridium tetani / enzymology*
Collagenases / antagonists & inhibitors,  chemistry*,  isolation & purification,  metabolism*
Escherichia coli / genetics
Gene Expression
Molecular Sequence Data
Phenanthrolines / pharmacology
Recombinant Proteins / antagonists & inhibitors,  chemistry,  isolation & purification,  metabolism
Reg. No./Substance:
0/Phenanthrolines; 0/Recombinant Proteins; 56-40-6/Glycine; 66-71-7/1,10-phenanthroline; 71-00-1/Histidine; EC 3.4.24.-/Collagenases

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