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Study of Receptor-Chaperone Interactions Using the Optical Technique of Spectroscopic Ellipsometry

By Verena Kriechbaumer, Anna Tsargorodskaya, Mohd K. Mustafa, Tatiana Vinogradova, Joanne Lacey, David P. Smith, Benjamin M. Abell and Alexei Nabok

Abstract

This work describes a detailed quantitative interaction study between the novel plastidial chaperone receptor OEP61 and isoforms of the chaperone types Hsp70 and Hsp90 using the optical method of total internal reflection ellipsometry (TIRE). The receptor OEP61 was electrostatically immobilized on a gold surface via an intermediate layer of polycations. The TIRE measurements allowed the evaluation of thickness changes in the adsorbed molecular layers as a result of chaperone binding to receptor proteins. Hsp70 chaperone isoforms but not Hsp90 were shown to be capable of binding OEP61. Dynamic TIRE measurements were carried out to evaluate the affinity constants of the above reactions and resulted in clear discrimination between specific and nonspecific binding of chaperones as well as differences in binding properties between the highly similar Hsp70 isoforms

Topics: Spectroscopy, Imaging, and Other Techniques
Publisher: The Biophysical Society
OAI identifier: oai:pubmedcentral.nih.gov:3136773
Provided by: PubMed Central
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