- Poster presentation
- Open access
- Published:
Impact of pro-domain stability of matrix metalloproteinase-8 on the outcome of sepsis
Critical Care volume 15, Article number: P278 (2011)
Introduction
Animal studies suggest matrix metalloproteinase-8 (MMP8) (neutrophil collagenase) impairs neutrophil (PMN) recruitment in inflammation; in humans, MMP8 has been associated with inflammation. We hypothesized that septic patients with single nucleotide polymorphisms (SNPs) in the MMP8 promoter region will have a survival advantage, and this advantage is due to differences in MMP8 enzymatic activity and not MMP8 levels.
Methods
We examined data from patients with CAP-associated sepsis (GenIMS), analyzed three functional SNPs (rs3765620, rs1940475, rs11225395) in 1,567 Caucasians and tested associations with 60-day and 90-day mortality and severe sepsis incidence. We simulated functional MMP8 SNPs using anisotropic network modeling. Modeling suggested pro-domain structural stability affecting zymogen activation. Based upon the predictions, we then studied zymogen activation using bioluminescent resonance energy transfer (BRET). We generated recombinant pro-MMP8 with a pro-domain tag of luciferase and carboxy terminus tag of green fluorescent protein. BRET signal was generated when luciferase-cleaved substrate produced a photon transferring energy to the GFP acceptor. GFP in turn emitted a green light signal when the donor/acceptor pairs were spatially close. Upon MMP activation, pro-domain is cleaved causing a loss in BRET signal.
Results
The rs1940475 genotype causing an amino acid mutation in the pro-domain was significantly associated with 90-day mortality (AA: 8.5%, AG: 11.1%, GG: 14.7%, P = 0.007). Cumulative incidence showed that the A allele was associated with better 90-day survival. Computer simulation of the mutation suggests a delayed activation. BRET assay confirmed that pro-domain mutation of MMP8 (K87E) rendered it less amenable to activation.
Conclusions
Our results suggest altering the structural stability of the inhibitory MMP8 pro-domain impacts enzyme activation. Therapeutics targeting pro-domain could be used to modulate MMP function and control downstream inflammatory processes in sepsis.
Author information
Authors and Affiliations
Rights and permissions
This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
About this article
Cite this article
McLaughlin, J., Rella, J., Bakan, A. et al. Impact of pro-domain stability of matrix metalloproteinase-8 on the outcome of sepsis. Crit Care 15 (Suppl 1), P278 (2011). https://doi.org/10.1186/cc9698
Published:
DOI: https://doi.org/10.1186/cc9698