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

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...

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Bibliographic Details
Published in:Photochemical & photobiological sciences 2015-10, Vol.14 (10), p.1844-1852
Main Authors: Tangorra, R. R., Operamolla, A., Milano, F., Hassan Omar, O., Henrard, J., Comparelli, R., Italiano, F., Agostiano, A., De Leo, V., Marotta, R., Falqui, A., Farinola, G. M., Trotta, M.
Format: Article
Language:English
Subjects:
Biochemistry
Biomaterials
Chemistry
Hydrophobic and Hydrophilic Interactions
Kinetics
Membranes, Artificial
Photosynthetic Reaction Center Complex Proteins - metabolism
Physical Chemistry
Plant Sciences
Polymers - chemistry
Protein Transport
Rhodobacter sphaeroides - enzymology
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Summary: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 PMOXA 22 -PDMS 61 -PMOXA 22 . 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 PMOXA 22 region of the polymersome, facing the water bulk solution, rather than in the PDMS 61 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.
ISSN:1474-905X
1474-9092
DOI:10.1039/c5pp00189g