Oriented immobilization of bacterial photosynthetic membrane

• Published in: Applied microbiology and biotechnology 1990-02, Vol.32 (5), p.544-549
• Main Authors: HARA, M, MAJIMA, T, MIYAKE, J, AJIKI, S, SUGINO, H, TOYOTAMA, H, KAWAMURA, S
• Format: Article
• Language: English
• Subjects: Adsorption; Antibodies, Bacterial - immunology; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; Bacteriological Techniques; Biological and medical sciences; Biotechnology; Biotin; Cell Membrane - physiology; Electrophoresis, Polyacrylamide Gel; Enzyme-Linked Immunosorbent Assay; Fundamental and applied biological sciences. Psychology; immobilization; Immobilization of organelles and whole cells; Immobilization techniques; Light; Membrane Potentials; membranes; Methods. Procedures. Technologies; photosynthesis; Photosynthesis - physiology; Photosynthetic Reaction Center Complex Proteins; Rhodopseudomonas - physiology
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/bf00173725

We have examined a method for oriented immobilization of photosynthetic membrane fragments on a solid surface by specific avidin-biotin interaction. Photosynthetic membrane fragments from the purple non-sulphur photosynthetic bacterium Rhodopseudomonas viridis, of which the H-subunit of the photosynthetic reaction centre was biotinylated, was immobilized on an avidin-adsorbed plate. Orientation of the immobilized membrane on the plastic plate was checked by an antisera binding assay that could react to the respective sides of the membrane: the H-subunit side was selectively adsorbed on the plate. Light-induced potential and current responses could be measured when the membrane immobilized on the SnO2-coated glass plate was dried and sandwiched with a counter electrode of Hg. The electrical response in the immobilized membrane was much improved in comparison with the control (membranes were simply adsorbed on the plate), supporting the idea that the membranes have an orientation on solid surfaces.