Direct visualization of protease action on collagen triple helical structure

Gabriel Rosenblum, Philippe E. van den Steen, Sidney R. Cohen, Arkady Bitler, David Brand, Ghislain Opdenakker, Irit Sagi

Research output: Contribution to journalArticle

42 Citations (Scopus)

Abstract

Enzymatic processing of extracellular matrix (ECM) macromolecules by matrix metalloproteases (MMPs) is crucial in mediating physiological and pathological cell processes. However, the molecular mechanisms leading to effective physiological enzyme-ECM interactions remain elusive. Only scant information is available on the mode by which matrix proteases degrade ECM substrates. An example is the enzymatic degradation of triple helical collagen II fragments, generated by the collagenase MMP-8 cleavage, during the course of acute inflammatory conditions by gelatinase B/MMP-9. As is the case for many other matrix proteases, it is not clear how MMP-9 recognizes, binds and digests collagen in this important physiological process. We used single molecule imaging to directly visualize this protease during its interaction with collagen fragments. We show that the initial binding is mediated by the diffusion of the protease along the ordered helix on the collagen 3/4 fragment, with preferential binding of the collagen tail. As the reaction progressed and prior to collagen degradation, gelatin-like morphologies resulting from the denaturation of the triple helical collagen were observed. Remarkably, this activity was independent of enzyme proteolysis and was accompanied by significant conformational changes of the working protease. Here we provide the first direct visualization of highly complex mechanisms of macromolecular interactions governing the enzymatic processing of ECM substrates by physiological protease.

Original languageEnglish (US)
Article numbere11043
JournalPloS one
Volume5
Issue number6
DOIs
StatePublished - Aug 10 2010

Fingerprint

collagen
Peptide Hydrolases
proteinases
Collagen
Visualization
Metalloproteases
metalloproteinases
extracellular matrix
Extracellular Matrix
Proteolysis
Physiological Phenomena
Macromolecular Substances
Degradation
acute course
Denaturation
gelatinase B
collagenase
degradation
Matrix Metalloproteinase 9
Collagenases

All Science Journal Classification (ASJC) codes

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

Rosenblum, G., van den Steen, P. E., Cohen, S. R., Bitler, A., Brand, D., Opdenakker, G., & Sagi, I. (2010). Direct visualization of protease action on collagen triple helical structure. PloS one, 5(6), [e11043]. https://doi.org/10.1371/journal.pone.0011043

Direct visualization of protease action on collagen triple helical structure. / Rosenblum, Gabriel; van den Steen, Philippe E.; Cohen, Sidney R.; Bitler, Arkady; Brand, David; Opdenakker, Ghislain; Sagi, Irit.

In: PloS one, Vol. 5, No. 6, e11043, 10.08.2010.

Research output: Contribution to journalArticle

Rosenblum, G, van den Steen, PE, Cohen, SR, Bitler, A, Brand, D, Opdenakker, G & Sagi, I 2010, 'Direct visualization of protease action on collagen triple helical structure', PloS one, vol. 5, no. 6, e11043. https://doi.org/10.1371/journal.pone.0011043
Rosenblum G, van den Steen PE, Cohen SR, Bitler A, Brand D, Opdenakker G et al. Direct visualization of protease action on collagen triple helical structure. PloS one. 2010 Aug 10;5(6). e11043. https://doi.org/10.1371/journal.pone.0011043
Rosenblum, Gabriel ; van den Steen, Philippe E. ; Cohen, Sidney R. ; Bitler, Arkady ; Brand, David ; Opdenakker, Ghislain ; Sagi, Irit. / Direct visualization of protease action on collagen triple helical structure. In: PloS one. 2010 ; Vol. 5, No. 6.
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