Cleavage of native type I collagen by human neutrophil elastase

被引:80
作者
Kafienah, W
Buttle, DJ
Burnett, D
Hollander, AP
机构
[1] Univ Sheffield, Sch Med, Dept Human Metab & Clin Biochem, Sheffield S10 2RX, S Yorkshire, England
[2] Univ Sheffield, Sch Med, Inst Bone & Joint Med, Sheffield S10 2RX, S Yorkshire, England
[3] Univ Birmingham, Micropathol Ltd, Inst Res & Dev, Birmingham B15 2SQ, W Midlands, England
关键词
D O I
10.1042/bj3300897
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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 degrees 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 alpha 2(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 mu g/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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页码:897 / 902
页数:6
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