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.

Assembly of a photosynthetic reaction center with ABA tri-block polymersomes : Highlights on protein localization / R.R. Tangorra, A. Operamolla, F. Milano, O.H. Omar, J. Henrard, R. Comparelli, F. Italiano, A. Agostiano, V. De Leo, R. Marotta, A. Falqui, G.M. Farinola, M. Trotta. - In: PHOTOCHEMICAL & PHOTOBIOLOGICAL SCIENCES. - ISSN 1474-905X. - 14:10(2015 Jul), pp. 1844-1852. [10.1039/c5pp00189g]

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

A. Falqui;
2015

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.
Settore FIS/03 - Fisica della Materia
Settore FIS/01 - Fisica Sperimentale
lug-2015
Article (author)
File in questo prodotto:
File Dimensione Formato  
Assembly of a photosynthetic reaction center_PHOTOCHEM&PHOTOBIOL Sci.pdf

accesso riservato

Descrizione: Articolo principale
Tipologia: Publisher's version/PDF
Dimensione 2.15 MB
Formato Adobe PDF
2.15 MB Adobe PDF   Visualizza/Apri   Richiedi una copia
Pubblicazioni consigliate

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/777401
Citazioni
  • ???jsp.display-item.citation.pmc??? 1
  • Scopus 15
  • ???jsp.display-item.citation.isi??? 15
social impact