Monomolecular Layer of the Alkylated Cytochrome b562 Formed at the Air/Water Interface

  • Kazutoshi Kobayashi
    Frontier Research Program, The Institute of Physical and Chemical Research (RIKEN)Hirosawa 2-1, Wako, Saitama 351-0198
  • Teruyuki Nagamune
    Department of Chemistry and Biotechnology, School of Engineering, The University of TokyoHongo 3-1, Bunkyo-ku, Tokyo 113-8656
  • Taiji Furuno
    School of Medicine, Keio UniversityHiyoshi 4-1-1, Kouhoku-ku, Yokohama, Kanagawa 223-8521
  • Hiroyuki Sasabe
    Frontier Research Program, The Institute of Physical and Chemical Research (RIKEN)Hirosawa 2-1, Wako, Saitama 351-0198

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<jats:title>Abstract</jats:title> <jats:p>A method is provided for forming a monomolecular layer of cytochrome b562 at the air/water interface by introducing dialkyl chains onto a water-soluble cytochrome b562. The purpose of the present study was to form densely and/or regularly packed two-dimensional arrays of protein molecules. Protoheme, which is a prosthetic group of cytochrome b562, was extracted from cytochrome b562, and then dialkylated(dioctadecylated) with the synthetic-chemical procedure. This dioctadecylated heme was reconstituted with apo-cytochrome b562 in dimethyl sulfoxide–water mixtures in vitro. The reconstitution reaction was successful when the reaction time was greater than 50 min, the dimethyl sulfoxide content was 63% and the temperature was maintained at 10 °C. Surface pressure–area per molecule isotherm of the reconstituted dioctadecylated holo-cytochrome b562 was measured and this clearly shows the monolayer formation of the water-soluble protein on the water subphase. The coefficient of compressibility of the protein film was estimated and is discussed. It is found that reconstituted dioctadecylated holo-cytochrome b562 forms a stable and closely packed monomolecular layer on the water surface.</jats:p>

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