Relationship between the individual collagenases of Clostridium histolyticum: evidence for evolution by gene duplication.
The relationship between the six collagenases (alpha, beta, gamma, delta, epsilon, and zeta) isolated and characterized in the preceding papers [Bond, M.D., & Van Wart, H.E. (1984) Biochemistry (preceding two papers in this issue)] has been investigated. Chemical modification reactions establish that all six enzymes contain essential carboxyl, tyrosine, and lysine residues. Circular dichroism spectra of the peptide bond region show that the secondary structures of the collagenases are very similar. Ouchterlony double-immunodiffusion experiments carried out with antiserum prepared against beta-collagenase indicate that all six collagenases are cross-reactive. Reverse-phase high-pressure liquid chromatography elution profiles of tryptic digests of these collagenases and sodium dodecyl sulfate electrophoresis gels of the peptides formed on reaction with cyanogen bromide have been obtained. The results indicate that the class I collagenases have extensive sequence homology with each other and that the class II collagenases have extensive sequence homology with each other but that the enzymes in the two classes have substantially different sequences. In addition, the data show that beta-collagenase probably consists of domains that have homologous amino acid sequences, which may have arisen by full or partial intragenic gene duplication. This may account for the unusually high molecular weight of this and the other collagenases. Finally, on the basis of the similarities between the collagenases in the two classes, it is suggested that one class evolved from the other by gene duplication followed by independent evolution by point mutations to yield enzymes with different substrate specificities.
D002942:Circular Dichroism; D003013:Clostridium / Q000201:enzymology* / Q000235:genetics; D005796:Genes; D005798:Genes, Bacterial; D007527:Isoenzymes / Q000235:genetics* / Q000378:metabolism; D003012:Microbial Collagenase / Q000235:genetics* / Q000378:metabolism; D011487:Protein Conformation
D007527:Isoenzymes; D003012:Microbial Collagenase