Acid Denaturation and Refolding of Cytochrome c on Silica Surface

• DOI: 10.1002/jccs.201200417
• Publication Date: 2012-11-19T13:31:38Z
• Authors: Christopher M. J....; Matthew T. Fontana; Benjamin C. Taggart; Meng‐Chih Su; Chung‐Lu Lin; Huan‐Cheng Chang; Hui‐Jung Chen
• Keywords: Denaturation (fissile materials); Protein Adsorption

Abstract Denaturation of oxidized cytochrome c (cyt c) adsorbed to a hydrophilic fused silica surface was studied by UV‐VIS attenuated total reflection (ATR) spectroscopy using multiple optical pass system newly developed this lab. Cyt adsorption at neutral pH gave an equilibrium constant K = 2 × 10 5 M −1 and coverage 63% monolayer saturation. Protein unfolding acid denaturation equilibrating bound cyt with buffers ranging in from 2. orientation were calculated based on theoretical model previous work. The average heme tilt angle (44°) found be independent pH, implicating protein‐surface interactions as the dominant factor governing adsorption. A non‐random molecular distribution observed, providing further support for dominance interactions. It shown that when denaturing removed replaced buffer refolded, assuming their original conformation. combination unique, yet applicable, science laboratory skills involved project had tremendous impact authors‘ undergraduate curriculum, making it ideal capstone development.