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Biochemistry Research InternationalVolume 2011 2011, Article ID 901572, 7 pages

Research ArticleBoston Biomedical Research Institute, 64 Grove Street, Watertown, MA 02472, USA

Received 29 April 2011; Accepted 11 July 2011

Academic Editor: Stefano Gianni

Copyright © 2011 Philip Graceffa. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.


Hsp27 oligomer is reported to interact with F-actin as a barbed-end-capping protein. The present study determined the binding strength and stoichiometry of the interaction using fluorescence of probes attached to Hsp27 cysteine-137. The fluorescence of acrylodan attached to Hsp27 increased 4-5-fold upon interaction with F-actin. Titration of the fluorescence with F-actin yielded a weak binding constant with an actin-Hsp27 stoichiometry between < 1 and 6. This stoichiometry is inconsistent with an F-actin end-capping protein. Pyrene attached to Hsp27 exhibited a large excimer fluorescence, in agreement with the known proximity of the cysteine-137-s in the Hsp27 oligomer. Upon interaction with F-actin the pyrene-Hsp27 excimer fluorescence was largely lost, suggesting that Hsp27 interacts with F-actin as a monomer, consistent with the acrylodan-Hsp27 results. EM images of F-actin-Hsp27 demonstrated that Hsp27 is not a strong G-actin sequester. Thus, Hsp27, in vitro, is a weak F-actin side-binding protein.

Author: Philip Graceffa

Source: https://www.hindawi.com/


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