Interferon gamma binds to extracellular matrix chondroitin-sulfate proteoglycans, thus enhancing its cellular...

The amino acid sequence of interferon gamma (IFN-gamma) has basic amino acid clusters similar to the heparin-binding consensus sequences found in other proteins that bind to proteoglycans (PGs). We investigated whether recombinant human IFN-gamma could bind to extracellular matrix (ECM) PGs secreted by human arterial smooth muscle cells (HASMCs) in vitro and whether the interaction affected the cellular response to IFN-gamma. As an in vitro model of ECM we used the basement membrane from HASMCs in culture. The binding of 125I-IFN-gamma to ECM was reduced significantly by pretreatment of ECM with chondroitinase ABC, an enzyme that degrades chondroitin-sulfate glycosaminoglycans. IFN-gamma binding to ECM was reduced by increasing concentrations of chondroitin-6-sulfate. 125I-IFN-gamma (0.05 to 2 ng/mL) binding data indicated an apparent Kd of 2 x 10(-11) mol/L and a maximum binding of 1.6 x 10(6) IFN-gamma molecules bound per square millimeter of ECM. Experiments with synthetic peptides suggested that residues 127 through 135 (AKTGKRKRS) are involved in the binding. The binding to chondroitin-sulfate PGs was confirmed by affinity chromatography of isolated [35S]chondroitin-sulfate PGs from ECM and cell-culture medium on immobilized IFN-gamma. The binding was abolished by treatment with chondroitinase ABC. ECM-bound IFN-gamma was more effective in inducing the expression of class II major histocompatibility antigens such as HLA-DR in HASMCs and human arterial endothelial cells than soluble IFN-gamma. These results suggest a role for chondroitin-sulfate PGs in immobilizing IFN-gamma in the ECM compartment and enhancing the cellular response to IFN-gamma.