Fabrication of ultra-thin 2D film of carbon nanotubes (CNTs) is achieved by employing a self-assembly protein of α-synuclein (αS). Electron energy loss spectroscopy-transmission electron microscopy and circular dichroism spectroscopy analyses indicate that αS binds to the hydrophobic surface of CNT through its N-terminal region by forming α-helix. ¹H-¹⁵N heteronuclear single-quantum coherence correlation-nuclear magnetic resonance spectra show that the acidic C-terminus of αS is exposed outward from the CNT surface. Following a slow vacuum filtration of the CNTs complexed with αS through a polycarbonate membrane filter, chloroform is used to unleash the free-floating ultra-thin CNT film from the substrate by causing unique αS-αS interactions of the enhanced anti-parallel β-sheet and α-helix formation. To make a graphically patterned CNT film stable in the free-floating state, gold nanoparticles (AuNPs) encapsulated with αS are employed to generate the αS-mediated CNT/AuNP hybrid film. The CNT-AuNP hybrid pattern is also directly transferred onto a thermo-responsive hydrogel sheet and sandwiched with a passive polymer layer. The resulting triply structure exhibits a shape deformation upon the light exposure. The light-responsive controllable soft material, therefore, has been suggested to be utilized in various areas including not only soft robotics but also human-machine interface by providing stimuli-responsive electroconductive and biocompatible matrix.