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Title: Structure of Protein Layers in Polyelectrolyte Matrices Studied by Neutron Reflectivity

Abstract

Polyelectrolyte multilayer films obtained by localized incorporation of Green Fluorescent Protein (GFP) within electrostatically assembled matrices of poly(styrene sulfonate)/poly(allylamine hydrochloride) (PSS/PAH) via spin-assisted layer-by-layer growth were discovered to be highly structured, with closely packed monomolecular layers of the protein within the bio-hybrid films. The structure of the films was evaluated in both vertical and lateral directions with neutron reflectometry, using deuterated GFP as a marker for neutron scattering contrast. Importantly, the GFP preserves its structural stability upon assembly as confirmed by circular dichroism (CD) and in situ attenuated total reflection Fourier Transform Infrared spectroscopy (ATR-FTIR). Atomic force microscopy was complimented with X-ray reflectometry to characterize the external roughness of the biohybrid films. Remarkably, films assembled with a single GFP layer confined at various distances from the substrate exhibit a strong localization of the GFP layer without intermixing into the LbL matrix. However, partial intermixing of the GFP layers with polymeric material is evidenced in multiple-GFP layer films with alternating protein-rich and protein-deficient regions. We hypothesize that the polymer-protein exchange observed in the multiple-GFP layer films suggests the existence of a critical protein concentration which can be accommodated by the multilayer matrix. Our results yield new insights into the mechanism ofmore » GFP interaction with a polyelectrolyte matrix and open opportunities for fabrication of bio-hybrid films with well-organized structure and controllable function, a crucial requirement for advanced sensing applications.« less

Authors:
 [1];  [2];  [2];  [2];  [1]
  1. University of Alabama, Birmingham
  2. ORNL
Publication Date:
Research Org.:
Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Spallation Neutron Source (SNS); Oak Ridge National Lab. (ORNL), Oak Ridge, TN (United States). Center for Structural Molecular Biology (CSMB)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1030620
DOE Contract Number:  
DE-AC05-00OR22725
Resource Type:
Journal Article
Journal Name:
Soft Matter
Additional Journal Information:
Journal Volume: 7; Journal Issue: 24; Journal ID: ISSN 1744-683X
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 99 GENERAL AND MISCELLANEOUS//MATHEMATICS, COMPUTING, AND INFORMATION SCIENCE; ATOMIC FORCE MICROSCOPY; DICHROISM; ELECTROLYTES; FABRICATION; FILMS; LAYERS; MATRICES; NEUTRON REFLECTORS; PROTEINS; REFLECTION; REFLECTIVITY; ROUGHNESS; SCATTERING; SPECTROSCOPY; SPIN; STABILITY; SUBSTRATES; Neutron Reflectivity; Layer-by-layer; Polyelectrolyte multilayer; Protein; Green Fluorescent Protein

Citation Formats

Kozlovskaya, Veronika, Ankner, John Francis, O'Neill, Hugh Michael, Zhang, Qiu, and Kharlampieva, Eugenia. Structure of Protein Layers in Polyelectrolyte Matrices Studied by Neutron Reflectivity. United States: N. p., 2011. Web. doi:10.1039/c1sm06710a.
Kozlovskaya, Veronika, Ankner, John Francis, O'Neill, Hugh Michael, Zhang, Qiu, & Kharlampieva, Eugenia. Structure of Protein Layers in Polyelectrolyte Matrices Studied by Neutron Reflectivity. United States. https://doi.org/10.1039/c1sm06710a
Kozlovskaya, Veronika, Ankner, John Francis, O'Neill, Hugh Michael, Zhang, Qiu, and Kharlampieva, Eugenia. 2011. "Structure of Protein Layers in Polyelectrolyte Matrices Studied by Neutron Reflectivity". United States. https://doi.org/10.1039/c1sm06710a.
@article{osti_1030620,
title = {Structure of Protein Layers in Polyelectrolyte Matrices Studied by Neutron Reflectivity},
author = {Kozlovskaya, Veronika and Ankner, John Francis and O'Neill, Hugh Michael and Zhang, Qiu and Kharlampieva, Eugenia},
abstractNote = {Polyelectrolyte multilayer films obtained by localized incorporation of Green Fluorescent Protein (GFP) within electrostatically assembled matrices of poly(styrene sulfonate)/poly(allylamine hydrochloride) (PSS/PAH) via spin-assisted layer-by-layer growth were discovered to be highly structured, with closely packed monomolecular layers of the protein within the bio-hybrid films. The structure of the films was evaluated in both vertical and lateral directions with neutron reflectometry, using deuterated GFP as a marker for neutron scattering contrast. Importantly, the GFP preserves its structural stability upon assembly as confirmed by circular dichroism (CD) and in situ attenuated total reflection Fourier Transform Infrared spectroscopy (ATR-FTIR). Atomic force microscopy was complimented with X-ray reflectometry to characterize the external roughness of the biohybrid films. Remarkably, films assembled with a single GFP layer confined at various distances from the substrate exhibit a strong localization of the GFP layer without intermixing into the LbL matrix. However, partial intermixing of the GFP layers with polymeric material is evidenced in multiple-GFP layer films with alternating protein-rich and protein-deficient regions. We hypothesize that the polymer-protein exchange observed in the multiple-GFP layer films suggests the existence of a critical protein concentration which can be accommodated by the multilayer matrix. Our results yield new insights into the mechanism of GFP interaction with a polyelectrolyte matrix and open opportunities for fabrication of bio-hybrid films with well-organized structure and controllable function, a crucial requirement for advanced sensing applications.},
doi = {10.1039/c1sm06710a},
url = {https://www.osti.gov/biblio/1030620}, journal = {Soft Matter},
issn = {1744-683X},
number = 24,
volume = 7,
place = {United States},
year = {Sat Jan 01 00:00:00 EST 2011},
month = {Sat Jan 01 00:00:00 EST 2011}
}