Cell surface chondroitin sulfate glycosaminoglycan in melanoma: role in the activation of pro-MMP-2 (pro-gelatinase A).

Abstract

We previously reported that CS (chondroitin sulfate) GAG (glycosaminoglycan), expressed on MCSP (melanoma-specific CS proteoglycan), is important for regulating MT3-MMP [membrane-type 3 MMP (matrix metalloproteinase)]-mediated human melanoma invasion and gelatinolytic activity in vitro. In the present study, we sought to determine if CS can directly enhance MT3-MMP-mediated activation of pro-MMP-2. Co-immunoprecipitation studies suggest that MCSP forms a complex with MT3-MMP and MMP-2 on melanoma cell surface. When melanoma cells were treated with betaDX (p-nitro-beta-D-xylopyranoside) to inhibit coupling of CS on the core protein, both active form and proform of MMP-2 were no longer co-immunoprecipitated with either MCSP or MT3-MMP, suggesting a model in which CS directly binds to MMP-2 and presents the gelatinase to MT3-MMP to be activated. By using recombinant proteins, we determined that MT3-MMP directly activates pro-MMP-2 and that this activation requires the interaction of the C-terminal domain of pro-MMP-2 with MT3-MMP. Activation of pro-MMP-2 by suboptimal concentrations of MT3-MMP is also significantly enhanced in the presence of excess C4S (chondroitin 4-sulfate), whereas C6S (chondroitin 6-sulfate) or low-molecular-mass hyaluronan was ineffective. Affinity chromatography studies using CS isolated from aggrecan indicate that the catalytic domain of MT3-MMP and the C-terminal domain of MMP-2 directly bind to the GAG. Thus the direct binding of pro-MMP-2 with CS through the C-domain would present the catalytic domain of pro-MMP-2 to MT3-MMP, which facilitates the generation of the active form of MMP-2. These results suggest that C4S, which is expressed on tumour cell surface, can function to bind to pro-MMP-2 and facilitate its activation by MT3-MMP-expressing tumour cells to enhance invasion and metastasis.

• Authors: Joji Iida, Krista L Wilhelmson, Janet Ng, Peter Lee, Charlotte Morrison, Eric Tam, Christopher M Overall, James B McCarthy
• Journal: The Biochemical journal (Biochem J) Vol. 403 Issue 3 Pg. 553-63 (May 01 2007) ISSN: 1470-8728 [Electronic] England
• PMID: 17217338 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• CSPG4 protein, human
• Chondroitin Sulfate Proteoglycans
• Enzyme Precursors
• Matrix Metalloproteinase Inhibitors
• Membrane Proteins
• Tissue Inhibitor of Metalloproteinase-2
• Chondroitin Sulfates
• Gelatinases
• Matrix Metalloproteinase 16
• Metalloendopeptidases
• progelatinase

Topics

• Chondroitin Sulfate Proteoglycans (metabolism)
• Chondroitin Sulfates (physiology)
• Enzyme Precursors (metabolism)
• Gelatinases (metabolism)
• Humans
• Matrix Metalloproteinase 16 (physiology)
• Matrix Metalloproteinase Inhibitors
• Melanoma (metabolism)
• Membrane Proteins (metabolism)
• Metalloendopeptidases (metabolism)
• Protein Structure, Tertiary
• Tissue Inhibitor of Metalloproteinase-2 (pharmacology)
• Tumor Cells, Cultured