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    How fluorescent labelling alters the solution behaviour of proteins


    Quinn, M. K. and Gnan, N. and James, S. and Ninarello, A. and Sciortino, F. and Zaccarelli, E. and McManus, Jennifer (2015) How fluorescent labelling alters the solution behaviour of proteins. Physical Chemistry Chemical Physics, 17. pp. 31177-31187. ISSN 1463-9076

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    Abstract

    A complete understanding of the role of molecular anisotropy in directing the self assembly of colloids and proteins remains a challenge for soft matter science and biophysics. For proteins in particular, the complexity of the surface at a molecular level poses a challenge for any theoretical and numerical description. A soft matter approach, based on patchy models, has been useful in describing protein phase behaviour. In this work we examine how chemical modification of the protein surface, by addition of a fluorophore, affects the physical properties of protein solutions. By using a carefully controlled experimental protein model (human gamma-D crystallin) and numerical simulations, we demonstrate that protein solution behaviour defined by anisotropic surface effects can be captured by a custom patchy particle model. In particular, the chemical modification is found to be equivalent to the addition of a large hydrophobic surface patch with a large attractive potential energy well, which produces a significant increase in the temperature at which liquid–liquid phase separation occurs, even for very low fractions of fluorescently labelled proteins. These results are therefore directly relevant to all applications based on the use of fluorescent labelling by chemical modification, which have become increasingly important in the understanding of biological processes and biophysical interactions.

    Item Type: Article
    Keywords: fluorescent labelling; solution behaviour; protein; molecular anisotropy; self assembly; colloids; proteins;
    Academic Unit: Faculty of Science and Engineering > Chemistry
    Item ID: 6631
    Identification Number: https://doi.org/10.1039/c5cp04463d
    Depositing User: Jennifer McManus
    Date Deposited: 01 Dec 2015 15:12
    Journal or Publication Title: Physical Chemistry Chemical Physics
    Publisher: Royal Society of Chemistry
    Refereed: Yes
    URI:
    Use Licence: This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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