Molecular Structures and Dynamics of the Stepwise Activation Mechanism of a Matrix Metalloproteinase Zymogen: Challenging the Cysteine Switch Dogma
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.
- Research Organization:
- Brookhaven National Lab. (BNL), Upton, NY (United States). National Synchrotron Light Source
- Sponsoring Organization:
- Doe - Office Of Science
- DOE Contract Number:
- DE-AC02-98CH10886
- OSTI ID:
- 959763
- Report Number(s):
- BNL-82749-2009-JA; JACSAT; TRN: US1005803
- Journal Information:
- Journal of the American Chemical Society, Vol. 129; ISSN 0002-7863
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
62 RADIOLOGY AND NUCLEAR MEDICINE
72 PHYSICS OF ELEMENTARY PARTICLES AND FIELDS
74 ATOMIC AND MOLECULAR PHYSICS
CYSTEINE
DISSOCIATION
ENZYMES
KALLIKREIN
MOLECULAR STRUCTURE
PROTEINS
PROTEOLYSIS
PROTONS
SYNERGISM
TRANSIENTS
WATER
X-RAY SPECTROSCOPY
ZINC IONS
national synchrotron light source