The recombinant catalytic domain of mouse collagenase-3 depolymerizes type I collagen by cleaving its aminotelopeptides.
The sequence coding for the catalytic domain of mouse collagenase-3 (MMP-13) was amplified by polymerase chain reaction and expressed in Escherichia coli. The recombinant catalytic domain (CCD), mainly recovered as inclusion bodies, was renatured and purified to homogeneity by preparative SDS-PAGE. The purified CCD degraded gelatin, casein and a synthetic peptide. CCD was not able to cleave the triple-helical domain of type I collagen but conserved the specific property of full-length collagenase-3 to cleave the N-telopeptides. These results show that residues involved in the recognition and cleavage of the aminotelopeptides of type I collagen are located in the catalytic domain of mouse collagenase-3 and that the C-terminal domain is not required for this activity.
Lemaître
V
V
Cell Biology Unit, University of Louvain Medical School, Bruxelles, Belgium.
Jungbluth
A
A
Eeckhout
Y
Y
D000818:Animals; D001665:Binding Sites; D003001:Cloning, Molecular; D003094:Collagen / Q000378:metabolism*; D017364:Collagenases / Q000378:metabolism*; D006168:Guinea Pigs; D053509:Matrix Metalloproteinase 13; D051379:Mice; D010446:Peptide Fragments / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D011994:Recombinant Proteins / Q000378:metabolism; D012886:Skull / Q000201:enzymology; D013379:Substrate Specificity
D010446:Peptide Fragments; D011994:Recombinant Proteins; D003094:Collagen; D017364:Collagenases; D053509:Matrix Metalloproteinase 13; C507081:Mmp13 protein, mouse