Strategies to fabricate biofunctionalized surfaces are essential for many biotechnological applications. Zwitterionic lipid bilayer coatings doped with lipids with chemically selective headgroups provide a robust platform for immobilization of biomolecules in an antifouling, protein resistant background. Herein, we assess the biological activity of two important components of the extracellular matrix (ECM), collagen type I (Col I) and fibronectin (FN), which are covalently attached to a supported lipid bilayer (SLB), and compare their activity with the same proteins, nonspecifically adsorbed onto a SiO2 surface. The characterization of protein coatings by quartz crystal microbalance with dissipation revealed that Col I and FN attached to SLB are less dense and have higher structural flexibility than when adsorbed onto SiO2. Cell adhesion, proliferation, and function, as well as Col I-FN interactions, were more efficient on the ECM-functionalized SLB, making it a promising platformfor cell-based diagnostics, tissue engineering, medical implants, and biosensor development.