Collagenase unwinds triple-helical collagen prior to peptide bond hydrolysis

被引：367

作者：

Linda Chung ^{[1
]}

Deendayal Dinakarpandian ^{[2
]}

Naoto Yoshida ^{[2
]}

Janelle L Lauer‐Fields ^{[2
]}

Gregg B Fields ^{[1
]}

Robert Visse ^{[3
]}

Hideaki Nagase ^{[1
]}

机构：

[1] Kennedy Inst. of Rheumatol. Division, Faculty of Medicine, Imperial College London, London

[2] Dept. of Biochem. and Molec. Biology, University of Kansas Medical Center, Kansas City, KS

[3] Dept. of Chemistry and Biochemistry, Florida Atlantic University, Boca Raton, FL

[4] Kennedy Inst. of Rheumatol. Division, Imperial College London, London W6 8LH

来源：

The EMBO Journal | 2004年 / 23卷 / 15期

关键词：

Collagenase; Enzyme mechanism; Matrix metalloproteinase; Protein unfolding; Triple helix;

D O I：

10.1038/sj.emboj.7600318

中图分类号：

学科分类号：

摘要：

Breakdown of triple-helical interstitial collagens is essential in embryonic development, organ morphogenesis and tissue remodelling and repair. Aberrant collagenolysis may result in diseases such as arthritis, cancer, atherosclerosis, aneurysm and fibrosis. In vertebrates, it is initiated by collagenases belonging to the matrix metalloproteinase (MMP) family. The three-dimensional structure of a prototypic collagenase, MMP-1, indicates that the substrate-binding site of the enzyme is too narrow to accommodate triple-helical collagen. Here we report that collagenases bind and locally unwind the triple-helical structure before hydrolyzing the peptide bonds. Mutation of the catalytically essential residue Glu200 of MMP-1 to Ala resulted in a catalytically inactive enzyme, but in its presence noncollagenolytic proteinases digested collagen into typical 3/4 and 1/4 fragments, indicating that the MMP-1 (E200A) mutant unwinds the triple-helical collagen. The study also shows that MMP-1 preferentially interacts with the α2(I) chain of type I collagen and cleaves the three α chains in succession. Our results throw light on the basic mechanisms that control a wide range of biological and pathological processes associated with tissue remodelling.

引用

页码：3020 / 3030

页数：10

共 54 条

[1]

Aimes R.T., Quigley J.P., Matrix metalloproteinase-2 is an interstitial collagenase. Inhibitor-free enzyme catalyzes the cleavage of collagen fibrils and soluble native type I collagen generating the specific 3/4- and 1/4-length fragments, J Biol Chem, 270, pp. 5872-5876, (1995)

[2]

Bode W., A helping hand for collagenases: The haemopexin-like domain, Structure, 3, pp. 527-530, (1995)

[3]

Brinckerhoff C.E., Matrisian L.M., Matrix metalloproteinases: A tail of a frog that became a prince, Nat Rev Mol Cell Biol, 3, pp. 207-214, (2002)

[4]

Brown R.A., Hukins D.W., Weiss J.B., Twose T.M., Do mammalian collagenases and DNA restriction endonucleases share a similar mechanism for cleavage site recognition?, Biochem Biophys Res Commun, 74, pp. 1102-1108, (1977)

[5]

Cawston T.E., Metalloproteinase inhibitors and the prevention of connective tissue breakdown, Pharmacol Therap, 70, pp. 163-182, (1996)

[6]

Chung L., Shimokawa K., Dinakarpandian D., Grams F., Fields G.B., Nagase H., Identification of the (RWTNNFREY191)-R-183 region as a critical segment of matrix metalloproteinase 1 for the expression of collagenolytic activity, J Biol Chem, 275, pp. 29610-29617, (2000)

[7]

Clark I.M., Cawston T.E., Fragments of human flbroblast collagenase. Purification and characterization, Biochem J, 263, pp. 201-206, (1989)

[8]

De Souza S.J., Pereira H.M., Jacchieri S., Brentani R.R., Collagen/collagenase interaction: Does the enzyme mimic the conformation of its own substrate?, FASEB J, 10, pp. 927-930, (1996)

[9]

Fields G.B., A model for interstitial collagen catabolism by mammalian collagenases, J Theor Biol, 153, pp. 585-602, (1991)

[10]

Fiori S., Sacca B., Moroder L., Structural properties of a collagenous heterotrimer that mimics the collagenase cleavage site of collagen type I, J Mol Biol, 319, pp. 1235-1242, (2002)

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