The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases

Role of the fibronectin type II-like repeats

J. Michael Shipley, Glenn A R Doyle, Catherine J. Fliszar, Qizhuang Ye, Linda L. Johnson, Steven D. Shapiro, Howard G. Welgus, Robert M. Senior

Research output: Contribution to journalArticle

158 Citations (Scopus)

Abstract

Several matrix metalloproteinases, including the 92-kDa and 72-kDa gelatinases, macrophage metalloelastase (MME), and matrilysin degrade insoluble elastin. Because elastolytieally active MME and matrilysin consist only of a catalytic domain (CD), we speculated that the homologous CDs of the 92-kDa and 72-kDa gelatinases would confer their elastolytic activities. In contrast to the MME CD, the 92 and 72 CDs expressed in Escherichia coli (lacking the internal fibronectin type II-like repeats) had no elastase activity, although both were gelatinolytic and cleaved a thiopeptolide substrate at rates comparable to the full-length gelatinases. To test the role of the fibronectin type II-like repeats in elastolytic activity, we expressed the 92-kDa gelatinase CD with its fibronectin type II-like repeats (92 CD/FN) in yeast. 92 CD/FN degraded insoluble elastin with activity comparable to full-length 92-kDa gelatinase. 92 and 72 CDs lacking the fibronectin type II-like repeats did not bind elastin, whereas the parent enzymes and 92 CD/FN did bind elastin. Furthermore, recombinant 92-kDa fibronectin type II-like repeats inhibited binding of the 92-kDa gelatinase to elastin. We conclude that the 92-and 72-kDa gelatinases require the fibronectin type II-like repeats for elastase activity.

Original languageEnglish (US)
Pages (from-to)4335-4341
Number of pages7
JournalJournal of Biological Chemistry
Volume271
Issue number8
StatePublished - Feb 23 1996
Externally publishedYes

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Matrix Metalloproteinase 2
Fibronectins
Elastin
Catalytic Domain
Matrix Metalloproteinase 12
Matrix Metalloproteinase 9
Matrix Metalloproteinase 7
Pancreatic Elastase
Gelatinases
Matrix Metalloproteinases
Yeast
Escherichia coli
Yeasts
Substrates
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Shipley, J. M., Doyle, G. A. R., Fliszar, C. J., Ye, Q., Johnson, L. L., Shapiro, S. D., ... Senior, R. M. (1996). The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases: Role of the fibronectin type II-like repeats. Journal of Biological Chemistry, 271(8), 4335-4341.

The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases : Role of the fibronectin type II-like repeats. / Shipley, J. Michael; Doyle, Glenn A R; Fliszar, Catherine J.; Ye, Qizhuang; Johnson, Linda L.; Shapiro, Steven D.; Welgus, Howard G.; Senior, Robert M.

In: Journal of Biological Chemistry, Vol. 271, No. 8, 23.02.1996, p. 4335-4341.

Research output: Contribution to journalArticle

Shipley, JM, Doyle, GAR, Fliszar, CJ, Ye, Q, Johnson, LL, Shapiro, SD, Welgus, HG & Senior, RM 1996, 'The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases: Role of the fibronectin type II-like repeats', Journal of Biological Chemistry, vol. 271, no. 8, pp. 4335-4341.
Shipley, J. Michael ; Doyle, Glenn A R ; Fliszar, Catherine J. ; Ye, Qizhuang ; Johnson, Linda L. ; Shapiro, Steven D. ; Welgus, Howard G. ; Senior, Robert M. / The structural basis for the elastolytic activity of the 92-kDa and 72-kDa gelatinases : Role of the fibronectin type II-like repeats. In: Journal of Biological Chemistry. 1996 ; Vol. 271, No. 8. pp. 4335-4341.
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