NDLI: Encapsulation into complex coacervate core micelles promotes EGFP dimerization

Content Provider	Royal Society of Chemistry (RSC)
Author	Westphal, A. H. Dongen, N. J. E. Berkel, W. J. H. Visser, A. J. W. G. Kleijn, J. M. Nolles, A. Borst, J. W.
Copyright Year	2017
Abstract	Complex coacervate core micelles (C3Ms) are colloidal structures useful for encapsulation of biomacromolecules. We previously demonstrated that enhanced green fluorescent protein (EGFP) can be encapsulated into C3Ms using the diblock copolymer poly(2-methyl-vinyl-pyridinium)41-b-poly(ethylene-oxide)205. This packaging resulted in deviating spectroscopic features of the encapsulated EGFP molecules. Here we show that for monomeric EGFP variant (mEGFP) micellar encapsulation affects the absorption and fluorescence properties to a much lesser extent, and that changes in circular dichroism characteristics are specific for encapsulated EGFP. Time-resolved fluorescence anisotropy of encapsulated (m)EGFP established the occurrence of homo-FRET (Förster resonance energy transfer) with larger transfer correlation times in the case of EGFP. Together, these findings support that EGFP dimerizes whereas the mEGFP mainly remains as a monomer in the densely packed C3Ms. We propose that dimerization of encapsulated EGFP causes a reorientation of Glu222, resulting in a pKa shift of the chromophore, which is fully reversible after release of EGFP from the C3Ms at a high ionic strength.
Starting Page	11380
Ending Page	11389
Page Count	10
File Format	HTM / HTML PDF
ISSN	14639076
Volume Number	19
Issue Number	18
Journal	Physical Chemistry Chemical Physics
DOI	10.1039/c7cp00755h
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Coacervate Förster resonance energy transfer Micelle Fluorescence Biomacromolecules EGFP Circular dichroism Anisotropy Ionic strength Green fluorescent protein Acid dissociation constant Copolymer Monomer Chromophore
Content Type	Text
Resource Type	Article
Subject	Physics and Astronomy Physical and Theoretical Chemistry

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