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Analytical technique of spectroscopic ellipsometry

Authors:
Alexei Nabok
Verena Kriechbaumer
Mohd Kamarulzaki Mustafa
Benjamin Abell
David Smith
Anna Tsargorodskaya

Keywords: chaperone, receptors, alpha-synuclein, spectroscopic ellipsometry, total internal reflection ellipsometry, electrostatic layer-by-layer deposition; AFM

Abstract:
The method of total spectroscopic ellipsometry in its total internal reflection (TIRE) mode was utilized in this work for the study of protein-protein interaction. Two different systems were studied here: (i) the interaction of chaperones, such as heat shock proteins Hsp70 and Hsp81 with different chaperone receptors including the new one OEP61-TM; (ii) the aggregation of alpha-synuclein monomers and oligomers (involved in Parkinson’s diseases) in phospholipid membranes. The method of TIRE provides unique quantified information on such protein interaction. Changes in the effective thickness of adsorbed molecular layer obtained by fitting the single TIRE spectra allows us to quantify the process of binding of chaperons to receptors or aggregation of alpha-synuclein in the lipid membrane. TIRE dynamic measurements allow monitoring protein interaction in-situ and evaluate the association (affinity) constants of such interaction. TIRE study revealed a clear separation between specific and non-specific interaction of proteins; such data were reported for the first time. A complementary method of atomic force microscopy (AFM) was used in this work to visualize the protein build-up on the surface.

Pages: 29 to 33

Copyright: Copyright (c) IARIA, 2011

Publication date: August 21, 2011

Published in: conference

ISSN: 2308-3514

ISBN: 978-1-61208-145-8

Location: Nice/Saint Laurent du Var, France

Dates: from August 21, 2011 to August 27, 2011