Hsp27-Actin Interaction
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...
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| Format: | Article |
| Language: | English |
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Wiley
2011-01-01
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| Series: | Biochemistry Research International |
| Online Access: | http://dx.doi.org/10.1155/2011/901572 |
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| _version_ | 1832548116451033088 |
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| author | Philip Graceffa |
| author_facet | Philip Graceffa |
| author_sort | Philip Graceffa |
| collection | DOAJ |
| description | 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 (KDapp=5.3 μM) 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. |
| format | Article |
| id | doaj-art-53f7ce853d834a7c83ccf7484c180ba7 |
| institution | Kabale University |
| issn | 2090-2247 2090-2255 |
| language | English |
| publishDate | 2011-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | Biochemistry Research International |
| spelling | doaj-art-53f7ce853d834a7c83ccf7484c180ba72025-02-03T06:42:02ZengWileyBiochemistry Research International2090-22472090-22552011-01-01201110.1155/2011/901572901572Hsp27-Actin InteractionPhilip Graceffa0Boston Biomedical Research Institute, 64 Grove Street, Watertown, MA 02472, USAHsp27 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 (KDapp=5.3 μM) 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.http://dx.doi.org/10.1155/2011/901572 |
| spellingShingle | Philip Graceffa Hsp27-Actin Interaction Biochemistry Research International |
| title | Hsp27-Actin Interaction |
| title_full | Hsp27-Actin Interaction |
| title_fullStr | Hsp27-Actin Interaction |
| title_full_unstemmed | Hsp27-Actin Interaction |
| title_short | Hsp27-Actin Interaction |
| title_sort | hsp27 actin interaction |
| url | http://dx.doi.org/10.1155/2011/901572 |
| work_keys_str_mv | AT philipgraceffa hsp27actininteraction |