TY - JOUR
T1 - Assembly of photosynthetic reaction center with ABA tri-block polymersomes: highlights on the protein localization.
AU - Tangorra, Roberto Rocco
AU - Operamolla, Alessandra
AU - Milano, Francesco
AU - Hassan Omar, Omar
AU - Henrard, John
AU - Comparelli, Roberto
AU - Italiano, Francesca
AU - Agostiano, Angela
AU - De Leo, Vincenzo
AU - Marotta, Roberto
AU - Falqui, Andrea
AU - Farinola, Gianluca
AU - Trotta, Massimo
N1 - KAUST Repository Item: Exported on 2020-10-01
PY - 2015
Y1 - 2015
N2 - 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 liposomes 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.
AB - 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 liposomes 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.
UR - http://hdl.handle.net/10754/559597
UR - http://pubs.rsc.org/en/Content/ArticleLanding/2015/PP/C5PP00189G
UR - http://www.scopus.com/inward/record.url?scp=84942863218&partnerID=8YFLogxK
U2 - 10.1039/C5PP00189G
DO - 10.1039/C5PP00189G
M3 - Article
SN - 1474-905X
VL - 14
SP - 1844
EP - 1852
JO - Photochem. Photobiol. Sci.
JF - Photochem. Photobiol. Sci.
IS - 10
ER -