Molecular structures and dynamics of the stepwise activation mechanism of a matrix metalloproteinase zymogen: Challenging the cysteine switch dogma

Gabriel Rosenblum, Samy Meroueh, Marta Toth, Jed F. Fisher, Rafael Fridman, Shahriar Mobashery, Irit Sagi

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

Activation of matrix metalloproteinase zymogen (pro-MMP) is a vital homeostatic process, yet its molecular basis remains unresolved. Using stopped-flow X-ray spectroscopy of the active site zinc ion, we determined the temporal sequence of pro-MMP-9 activation catalyzed by tissue kallikrein protease in milliseconds to several minutes. The identity of three intermediates seen by X-ray spectroscopy was corroborated by molecular dynamics simulations and quantum mechanics/molecular mechanics calculations. The cysteine-zinc interaction that maintains enzyme latency is disrupted via active-site proton transfers that mediate transient metal-protein coordination events and eventual binding of water. Unexpectedly, these events ensue as a direct result of complexation of pro-MMP-9 and kallikrein and occur before proteolysis and eventual dissociation of the pro-peptide from the catalytic site. Here we demonstrate the synergism among long-range protein conformational transitions, local structural rearrangements, and fine atomic events in the process of zymogen activation.

Original languageEnglish (US)
Pages (from-to)13566-13574
Number of pages9
JournalJournal of the American Chemical Society
Volume129
Issue number44
DOIs
StatePublished - Nov 7 2007
Externally publishedYes

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Enzyme Precursors
Molecular Dynamics Simulation
Molecular Structure
Matrix Metalloproteinases
Molecular structure
Cysteine
Molecular dynamics
Chemical activation
Switches
X ray spectroscopy
Catalytic Domain
Zinc
Mechanics
Proteolysis
Proteins
Spectrum Analysis
Molecular mechanics
Proton transfer
Quantum theory
X-Rays

ASJC Scopus subject areas

  • Chemistry(all)

Cite this

Molecular structures and dynamics of the stepwise activation mechanism of a matrix metalloproteinase zymogen : Challenging the cysteine switch dogma. / Rosenblum, Gabriel; Meroueh, Samy; Toth, Marta; Fisher, Jed F.; Fridman, Rafael; Mobashery, Shahriar; Sagi, Irit.

In: Journal of the American Chemical Society, Vol. 129, No. 44, 07.11.2007, p. 13566-13574.

Research output: Contribution to journalArticle

Rosenblum, Gabriel ; Meroueh, Samy ; Toth, Marta ; Fisher, Jed F. ; Fridman, Rafael ; Mobashery, Shahriar ; Sagi, Irit. / Molecular structures and dynamics of the stepwise activation mechanism of a matrix metalloproteinase zymogen : Challenging the cysteine switch dogma. In: Journal of the American Chemical Society. 2007 ; Vol. 129, No. 44. pp. 13566-13574.
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