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Matrix metalloproteinase 9 (MMP-9)-dependent processing of βig-h3 protein regulates cell migration, invasion, and

Yeon Hyang Kim1, Hyung-Joo Kwon, Doo-Sik Kim

  • 1Department of Biochemistry, College of Life Science and Biotechnology, Yonsei University, Seoul 120-749, Korea. yeonhkim@yonsei.ac.kr

The Journal of Biological Chemistry
|September 29, 2012
PubMed

Insights

Matrix metalloproteinase-9 (MMP-9) cleaves transforming growth factor-β-induced protein (βig-h3), altering cell migration and extracellular matrix interactions. This cleavage promotes tumor cell and macrophage invasion and migration.

Area of Science:

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Background:

  • Cell migration is crucial for biological processes like inflammation, cancer, and development.
  • Transforming growth factor-β-induced protein (βig-h3) is implicated in cellular functions.
  • Matrix metalloproteinase-9 (MMP-9) is a key enzyme in extracellular matrix remodeling.

Purpose of the Study:

  • To investigate the interaction between βig-h3 and MMP-9.
  • To determine the functional consequences of MMP-9-mediated βig-h3 cleavage on cell behavior.
  • To elucidate the role of βig-h3 and its cleavage products in cell migration and invasion.

Main Methods:

  • Site-directed mutagenesis to identify MMP-9 cleavage sites on βig-h3.
  • Expression and purification of recombinant βig-h3 and MMP-9.
  • Proteolytic cleavage assays using various cell lines (HEK293F, U87MG) and primary cells (macrophages, neutrophils).
  • Analysis of cell adhesion, cell-cell interactions (spheroid formation), and cell invasion.
  • Macrophage migration assays and signaling pathway analysis (FAK/Src).

Main Results:

  • MMP-9 proteolytically cleaves βig-h3 at specific sites.
  • Cleavage releases βig-h3 from extracellular matrix proteins (collagen IV, fibronectin) and abolishes its adhesive properties.
  • MMP-9-mediated βig-h3 cleavage increases glioma cell invasion.
  • Knockdown of βig-h3 also enhances cell invasion.
  • Cleaved βig-h3 fragments can bind to macrophages and act as chemoattractants, inducing migration via FAK/Src signaling.
  • Intact βig-h3 inhibits cell migration, promotes cell-cell contact, and cell-ECM interaction.

Conclusions:

  • MMP-9 cleavage of βig-h3 significantly alters its function, promoting cell migration and invasion.
  • Cleaved βig-h3 acts as a chemoattractant for macrophages, contributing to MMP-9-driven tumor cell and macrophage migration.
  • βig-h3 plays opposing roles in cell behavior depending on its cleavage state, with intact βig-h3 inhibiting migration and cleaved βig-h3 promoting it.

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