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Assembly of a photosynthetic reaction center with ABA tri-block polymersomes: highlights on protein localization

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Abstract

The reconstitution of the integral membrane protein photosynthetic reaction center (RC) in polymersomes, i.e. artificial closed vesicles, was achieved by the micelle-to-vesicle transition technique, a very mild protocol based on size exclusion chromatography often used to drive the incorporation of proteins contemporarily to liposome formation. An optimized protocol was used to successfully reconstitute the protein in a fully active state in polymersomes formed by the tri-block copolymers PMOXA22-PDMS61-PMOXA22. The RC is very sensitive to its solubilizing environment and was used to probe the positioning of the protein in the vesicles. According to charge-recombination experiments and to the enzymatic activity assay, the RC is found to accommodate in the PMOXA22 region of the polymersome, facing the water bulk solution, rather than in the PDMS61 transmembrane-like region. Furthermore, polymersomes were found to preserve protein integrity efficiently as the biomimetic lipid bilayers but show a much longer temporal stability than lipid based vesicles.

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Authors and Affiliations

  1. Chemistry Department, Università degli Studi di Bari “Aldo Moro”, via Orabona 4, 70125, Bari, Italy

    R. R. Tangorra, A. Operamolla, A. Agostiano, V. De Leo & G. M. Farinola

  2. CNR-IPCF, Istituto per i processi Chimico-Fisici, UOS Bari (Italy), via Orabona 4, 70125, Bari, Italy

    F. Milano, R. Comparelli, F. Italiano, A. Agostiano & M. Trotta

  3. CNR-ICCOM, Istituto di Chimica dei Composti Organometallici, via Orabona 4, 70125, Bari, Italy

    O. Hassan Omar & G. M. Farinola

  4. Chemistry Department and Namur Research College (NARC), University of Namur (UNamur), Rue de Bruxelles 61, Namur, Belgium

    J. Henrard

  5. Electron Microscopy Lab, Nanochemistry Department Istituto Italiano di Tecnologia (IIT), Via Morego 30, 16163, Genova, Italy

    R. Marotta

  6. King Abdullah University of Science and Technology, Biological and Environmental Sciences and Engineering Division, 23955-6900, Thuwal, Saudi Arabia

    A. Falqui

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  1. R. R. Tangorra
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  2. A. Operamolla
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  3. F. Milano
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  4. O. Hassan Omar
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  6. R. Comparelli
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  7. F. Italiano
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  8. A. Agostiano
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  9. V. De Leo
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  10. R. Marotta
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  11. A. Falqui
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  12. G. M. Farinola
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  13. M. Trotta
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Corresponding author

Correspondence to G. M. Farinola.

Additional information

Electronic supplementary information (ESI) available: Synthesis details of ABA and supplementary figures. See DOI: 10.1039/c5pp00189g

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Tangorra, R.R., Operamolla, A., Milano, F. et al. Assembly of a photosynthetic reaction center with ABA tri-block polymersomes: highlights on protein localization. Photochem Photobiol Sci 14, 1844–1852 (2015). https://doi.org/10.1039/c5pp00189g

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  • DOI: https://doi.org/10.1039/c5pp00189g

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