Proteomic analysis identifies in vivo candidate matrix metalloproteinase-9 substrates in the left ventricle post-myocardial infarction

Rogelio Zamilpa, Elizabeth F. Lopez, Ying Ann Chiao, Qiuxia Dai, Gladys P. Escobar, Kevin Hakala, Susan T. Weintraub, Merry L. Lindsey

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68 Scopus citations

Abstract

Matrix metalloproteinase-9 (MMP-9) deletion has been shown to improve remodeling of the left ventricle post-myocardial infarction (MI), but the mechanisms to explain this improvement have not been fully elucidated. MMP-9 has a broad range of in vitro substrates, but relevant in vivo substrates are incompletely defined. Accordingly, we evaluated the infarct regions of wild-type (wt) and MMP-9 null (null) mice using a proteomic strategy. Wt and null groups showed similar infarct sizes (48±3 in wt and 45±3% in null), indicating that both groups received an equal injury stimulus. Left ventricle infarct tissue was homogenized and analyzed by 2-DE and MS. Of 31 spot intensity differences, the intensities of 9 spots were higher and 22 spots were lower in null mice compared to wt (all p<0.05). Several extracellular matrix proteins were identified in these spots by MS, including fibronectin, tenascin-C, thrombospondin-1, and laminin. Fibronectin was observed on the gels at a lower than expected molecular weight in the wt group, which suggested substrate cleavage, and the lower molecular weight spot was observed at lower intensity in the MMP-9 null group, which suggested cleavage by MMP-9. Immunoblotting confirmed the presence of fibronectin cleavage products in the wt samples and lower levels in the absence of MMP-9. In conclusion, examining infarct tissue from wt and MMP-9 null mice by proteomic analysis provides a powerful and unique method to identify in vivo candidate MMP substrates.

Original languageEnglish (US)
Pages (from-to)2214-2223
Number of pages10
JournalProteomics
Volume10
Issue number11
DOIs
Publication statusPublished - Jun 1 2010

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Keywords

  • Animal proteomics
  • Cardiac remodeling
  • Extracellular matrix
  • MMP-9
  • Mice
  • Myocardial infarction

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

Cite this

Zamilpa, R., Lopez, E. F., Chiao, Y. A., Dai, Q., Escobar, G. P., Hakala, K., ... Lindsey, M. L. (2010). Proteomic analysis identifies in vivo candidate matrix metalloproteinase-9 substrates in the left ventricle post-myocardial infarction. Proteomics, 10(11), 2214-2223. https://doi.org/10.1002/pmic.200900587