title: Building a novel vitronectin assay by immobilization of integrin on calixarene monolayer.
abstract: Membrane proteins possess significant hydrophobic domains and are likely to deplete their native activity immobilized on the solid surface relative to those occurring in a membrane environment. To investigate an efficient immobilization method, calix[4]crown-ether monolayer as an artificial protein linker system was constructed on the gold surface and characterized by Fourier transform infrared reflection absorption spectroscopy (FTIR-RAS), atomic force microscopy (AFM) and cyclic voltammetry (CV). Integrin alpha(v)beta3 was functionally immobilized onto the monolayer and the integrin-vitronectin interaction was investigated by surface plasmon resonance (SPR). It was found that calix[4]crown-ether was assembled as a monolayer on the gold surface. Orientation and accessibility of integrin alpha(v)beta3 was assessed by sensitive binding of its natural ligand, vitronectin at pg mL(-1) level. Moreover, surface coverage of integrin layer and thickness calculated through SPR curve simulation verified that integrin layer was a monolayer in activated form. In combination with the SPR method, this calix[4]crown monolayer provided a reliable and simple experimental platform for the investigation of isolated membrane proteins under experimental conditions resembling those of their native properties.
authors: College of Pharmacy, Pusan National University, Pusan 609-735, Republic of Korea.
authors: Hongxia
authors: H
authors: Chen
authors: Minsu
authors: M
authors: Lee
authors: Jaebeom
authors: J
authors: Lee
authors: Won Gun
authors: WG
authors: An
authors: Heung-Jin
authors: HJ
authors: Choi
authors: Sung-Hoon
authors: SH
authors: Kim
authors: Kwangnak
authors: K
authors: Koh
mesh_terms: D047250:Calixarenes / Q000737:chemistry*; D004563:Electrochemistry; D039302:Integrin alphaVbeta3 / Q000737:chemistry*; D018625:Microscopy, Atomic Force; D015394:Molecular Structure; D017550:Spectroscopy, Fourier Transform Infrared; D020349:Surface Plasmon Resonance / Q000379:methods; D019096:Vitronectin / Q000737:chemistry*
chemical_list: D039302:Integrin alphaVbeta3; D019096:Vitronectin; D047250:Calixarenes