Cleavage of native type I collagen by human neutrophil elastase

W KAFIENAH, JD BUTTLE, D BURNETT… - Biochemical …, 1998 - portlandpress.com
W KAFIENAH, JD BUTTLE, D BURNETT, PA HOLLANDER
Biochemical Journal, 1998portlandpress.com
The ability of purified human neutrophil elastase (EC 3.4. 21.37) to cleave native type I
collagen has been investigated. Soluble human, bovine or rat type I collagen was incubated
with neutrophil elastase for 16 h at 25° C before catalysis was stopped with 3, 4-
dichloroisocoumarin. Analysis by SDS/PAGE of the collagen digests revealed 3/4-length
fragments similar in size to those produced by interstitial collagenase. The collagenolytic
activity was dose dependent and was not due to a contaminating metalloproteinase or …
The ability of purified human neutrophil elastase (EC 3.4.21.37) to cleave native type I collagen has been investigated. Soluble human, bovine or rat type I collagen was incubated with neutrophil elastase for 16 h at 25 °C before catalysis was stopped with 3,4-dichloroisocoumarin. Analysis by SDS/PAGE of the collagen digests revealed 3/4-length fragments similar in size to those produced by interstitial collagenase. The collagenolytic activity was dose dependent and was not due to a contaminating metalloproteinase or cysteine proteinase, as it was not inhibited by 1,10-phenanthroline, EDTA or L-trans-epoxysuccinyl-leucylamido-(4-guanidino)butane. The identity of the cleavage products was confirmed using a new antibody that recognizes the unwound α2(I)-chain. This detected the 3/4-length fragment of type I collagen following neutrophil elastase cleavage. In addition to cleaving soluble collagen, neutrophil elastase also cleaved reconstituted, radiolabelled type I collagen fibrils, at a rate of 16 μg/min per nmol. These results indicate that neutrophil elastase can cleave native type I collagen in the helix, an activity that might contribute to its roles in connective-tissue pathology.
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